http://2010.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=50&target=Djsexton5&year=&month=2010.igem.org - User contributions [en]2024-03-28T22:58:09ZFrom 2010.igem.orgMediaWiki 1.16.5http://2010.igem.org/Team:Georgia_State/OursystemTeam:Georgia State/Oursystem2010-10-28T03:21:59Z<p>Djsexton5: /* H1N1 VACCINE */</p>
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<div>{{Georgia_State/Header}}<br />
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== H1N1 VACCINE ==<br />
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'''Background''':<br />
In 2009 the World Health Organization raised its pandemic alert for H1N1 influenza A <br />
virus to the 6 phase alert. This is the highest alert level and it indicates widespread<br />
community transmission of the virus in over two continents. The H1N1 Influenza A virus<br />
is a quadruple reassortment of two swine strains, one human strain, and one avian strain.<br />
More than 214 countries and territories have reported laboratory – confirmed cases of<br />
pandemic H1N1 Influenza A. The US Centers for Disease Control and Prevention reported<br />
between April 2009 and April 2010, approximately 61million cases of pandemic H1N1 occurred,<br />
including 274,000 hospitalizations and 12,470 deaths. <br />
<br />
'''Problem:''' <br />
Efficient and timely vaccine production was a challenge for the scientific<br />
community during the pandemic outbreak of H1N1. Another problem with the <br />
Influenza viruses is the high variability and mutations that they present<br />
with each year. This makes vaccine production time difficult and time consuming.<br />
<br />
'''The Pichia System:'''<br />
The Pichia system is an ideal construct for producing vaccines. We have designed a standard Pichia system that can be used for the production of various different vaccines. In theory, a high yield circuit can be established that only requires the exchange of the variable antigen, while leaving the remaining factors constant. In the circuit presented below, a vector has been designed to express the HA globular head influenza A antigen under the control of the strong constitutive promter pGAP. <br />
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[[Image:H1N1.png|Center|atl=Alt text|H1N1 INFLUENZA A VIRUS]]<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_StateTeam:Georgia State2010-10-28T03:01:42Z<p>Djsexton5: </p>
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[[Image:GSUteamphoto.jpg|900px|center]]<br />
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==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|left|200px]]<br />
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''Pichia pastoris'' is a methylotrophic yeast used as an alternative host for protein production in addition to ''Escherichia coli'' and ''Saccharomyces cerevisiae''. There are several reasons why ''P. Pastoris'' is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
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Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer![[Image:Pichia_tool_box_picture.jpg|right]]<br />
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== '''Check out the current toolbox here!!''' == [[Team:Georgia State/Parts]]<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/WhyPichiaTeam:Georgia State/WhyPichia2010-10-28T02:54:28Z<p>Djsexton5: /* Pichia Vs Saccharomyces */</p>
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<div>{{Georgia_State/Header}}<br />
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[[Image:Whypichia.png|500px|right]]<br />
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=='''Pichia Vs Saccharomyces'''== <br />
<br />
<br />
'''Greater Cell Concentrations'''<br />
The preference of Pichia pastoris for respiratory growth is a key characteristic which allows it to be cultured at high cell densities (500 OD600 U ml31)[1]. This trait lends an advantage to Pichia over Saccharomyces cervisiae: Resultant ethanol from S. cervisiae fermentation accumulates which hinders culture growth and therefore protein production. High levels of cell growth in fermenter cultures are vital to foreign protein production since concentration of the secreted product in "extracellular medium is...proportional to the concentration of cells in the culture". [2] <br />
<br />
General notworthy differences inlcude the relatively stable ell membrane lipids of Pichia that form etter biocatalyst.(jiang, 2008). Pichia also has more compact and organized golgi bodies than Saccharomyces(Morawski, 2000). In contrast, there are many similarities between Pichia and Saccharomyces that greatly increase the benefits affiliated with using Pichia. Genetic similarity between the two organisms has enabled expression of similar genes and compatibility between vectors. Other similarities include identical pathways for TCA, pentose phosphate, glycolysis and amino acid synthesis(Fiaux, 2003) and (Baumann, 2010).<br />
<br />
=='''Easy and Inexpensive to Culture'''==<br />
<br />
<br />
The components of P. pastoris media (glycerol, methanol, salts, trace elements, and biotin) are relatively inexpensive and as such are well suited for large-scale heterologous protein production. The common protocol is to allow P. pastoris to accumulate biomass in a glycerol/carbon-source medium while foreign protein expression is repressed. Once glycerol stocks have been depleted, methanol is added to induce expression. Any undesirable protease activity can be easily averted by adjusting pH. Pichia's broad spectrum of optimal growth pH (3.0-7.0) provides ample wiggle room. A number of other techniques may be employed in minimizing proteolysis including lowering growth rate via methanol-level modulation, addition of protease inhibitors, addition of alternative protease substrates, and even simply lowering process temperature.<br />
<br />
<br />
== '''High production of foreign Protein while low levels of endogenous protein''' ==<br />
<br />
<br />
Extracellular expression of foreign proteins is favorable to intracellular production due to Pichia's extremely low levels of endogenous protein secretion, which makes it easier to isolate the protein of interest. Pichia vectors can also be cloned with foreign genes which align with a secretion signal (native to the protein, native to Pichia, or native to Saccharomyces), facilitating extracellular production. However, intracellular protein production is also high due to the efficiency--at carefully monitored methanol levels--of the AOX promoter. <br />
<br />
<br />
[1] Cregg 1999 review paper<br />
<br />
[2] Heterologous protein production using the Pichia pastoris expression system (2005). Sue Macaulet-Patric, mariana L. Fazenda, Brian McNeil, and Linda M. Harvey. Yeast (22): 249-270<br />
<br />
<br />
<br />
== '''Pichia pastoris versus Esherichia coli''' ==<br />
<br />
<br />
The bacterium E. coli has been frequently chosen as a host to express recombinant, heterologous proteins. Although many successful experiments have used E. coli as their expression system, P. pastoris has become an increasingly popular alternative due to the shortcomings of E. coli. P. pastoris serves as a better host when attempting to express foreign genes because this yeast can perform post-translational modifications and fold proteins properly, tightly regulate transcription via its wide range of promotors, and be purified easily.<br />
<br />
a. Post-translational modifications<br />
<br />
Unlike P. pastoris, E. coli cannot perform higher eukaryotic post-translational modifications. As a result, proteins requiring these modifications often fold incorrectly when produced by E. coli. These disadvantages, which result from E. coli’s prokaryotic nature, limit the types of proteins that this system can express. Proteins that contain disulfide bonds or require post-translational modifications such as glycosylation, isomerization, or phosphorylation are not always properly expressed; they can be insoluble or improperly folded, which require additional solubilization and re-folding steps (Daly et al). Take for instance erythropoietin (EPO), a glycosylated protein. When EPO is expressed in E. coli, it is not glycosylated and becomes less resistant to unfolding compared to its natural, glycosylated form (Daly et al). Therefore, additional steps are needed in order to form a stable protein, which is time-consuming, costly, and has the tendency to produce low yields. <br />
<br />
A study done by Leuking et al. also supports the use of P. pastoris for protein expression. Vectors were created for both E. coli and P. pastoris using multiple cDNAs from a human fetal brain expression library. Out of the 29 DNA clones, all produced soluble proteins in P. pastoris, while E. coli was much less successful. In E. coli, only nine produced soluble proteins; 15 were detected as inclusion bodies; and five were not expressed at all. These differences are likely due to E. coli’s lack of eukaryotic abilities to perform post-translational modifications and properly fold proteins (Leuking et al).<br />
<br />
b. Powerful promoter systems<br />
<br />
P. pastoris’s wide range of promotors also aid in its use for foreign protein production. It has many strongly induced promotors, which allow sufficient expression of the genes of interest and therefore, a high concentration of proteins can be easily and inexpensively produced (Weidner et al). The alcohol oxidase genes, AOX1 and AOX2, are frequently used promotors because they are easily induced by methanol; because P. pastoris is methylotropic, meaning it can metabolize methanol as its carbon and energy source, adding methanol to the growth medium will induce these two promotors and then express the desired protein (Macauley-Patrick et al). High concentrations of protein can then be produced.<br />
A third advantage that P. pastoris has over E. coli is that proteins expressed by P. pastoris are easily purified (Weidner et al). Because P. pastoris secretes its recombinant protein in the growth medium and secretes low levels of endogeneous proteins, there is a better chance of obtaining high yields of uncontaminated proteins in comparison to E. coli (Weidner et al). In essence, there are fewer steps required to obtain the desired protein.<br />
<br />
<br />
Works Cited<br />
Daly, Rachel, and Milton T. W. Hearn. "Expression of Heterologous Proteins in Pichia pastoris: a Useful Experimental Tool in Protein Engineering and Production." PubMed.gov. PubMed, 26 Nov. 2004. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15565717>.<br />
Leuking, Angelika, Caterina Holz, Christine Gotthold, Hans Lehrach, and Dolores Cahill. “A System for Dual Protein Expression in Pichia pastoris and Escherichia coli.” PubMed.gov. PubMed, 15 Dec. 2003. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
Macauley-Patrick, Sue, Mariana L. Fazenda, Brian McNeil, and Linda M. "Heterologous Protein Production Using the Pichia pastoris Expression System." PubMed.gov. PubMed, 22 Mar. 2005. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15704221>.<br />
Weidner, Maria, Marcus Taupp, and Steven J. Hallam. "Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris." PubMed.gov. PubMed, 25 Feb. 2010. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/WhyPichiaTeam:Georgia State/WhyPichia2010-10-28T02:43:11Z<p>Djsexton5: /* Pichia Vs Saccharomyces */</p>
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<div>{{Georgia_State/Header}}<br />
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[[Image:Whypichia.png|500px|right]]<br />
<br />
<br />
=='''Pichia Vs Saccharomyces'''== <br />
<br />
<br />
'''Greater Cell Concentrations'''<br />
The preference of Pichia pastoris for respiratory growth is a key characteristic which allows it to be cultured at high cell densities (500 OD600 U ml31)[1]. This trait lends an advantage to Pichia over Saccharomyces cervisiae: Resultant ethanol from S. cervisiae fermentation accumulates which hinders culture growth and therefore protein production. High levels of cell growth in fermenter cultures are vital to foreign protein production since concentration of the secreted product in "extracellular medium is...proportional to the concentration of cells in the culture". [2]<br />
<br />
=='''Easy and Inexpensive to Culture'''==<br />
<br />
<br />
The components of P. pastoris media (glycerol, methanol, salts, trace elements, and biotin) are relatively inexpensive and as such are well suited for large-scale heterologous protein production. The common protocol is to allow P. pastoris to accumulate biomass in a glycerol/carbon-source medium while foreign protein expression is repressed. Once glycerol stocks have been depleted, methanol is added to induce expression. Any undesirable protease activity can be easily averted by adjusting pH. Pichia's broad spectrum of optimal growth pH (3.0-7.0) provides ample wiggle room. A number of other techniques may be employed in minimizing proteolysis including lowering growth rate via methanol-level modulation, addition of protease inhibitors, addition of alternative protease substrates, and even simply lowering process temperature.<br />
<br />
<br />
== '''High production of foreign Protein while low levels of endogenous protein''' ==<br />
<br />
<br />
Extracellular expression of foreign proteins is favorable to intracellular production due to Pichia's extremely low levels of endogenous protein secretion, which makes it easier to isolate the protein of interest. Pichia vectors can also be cloned with foreign genes which align with a secretion signal (native to the protein, native to Pichia, or native to Saccharomyces), facilitating extracellular production. However, intracellular protein production is also high due to the efficiency--at carefully monitored methanol levels--of the AOX promoter. <br />
<br />
<br />
[1] Cregg 1999 review paper<br />
<br />
[2] Heterologous protein production using the Pichia pastoris expression system (2005). Sue Macaulet-Patric, mariana L. Fazenda, Brian McNeil, and Linda M. Harvey. Yeast (22): 249-270<br />
<br />
<br />
<br />
== '''Pichia pastoris versus Esherichia coli''' ==<br />
<br />
<br />
The bacterium E. coli has been frequently chosen as a host to express recombinant, heterologous proteins. Although many successful experiments have used E. coli as their expression system, P. pastoris has become an increasingly popular alternative due to the shortcomings of E. coli. P. pastoris serves as a better host when attempting to express foreign genes because this yeast can perform post-translational modifications and fold proteins properly, tightly regulate transcription via its wide range of promotors, and be purified easily.<br />
<br />
a. Post-translational modifications<br />
<br />
Unlike P. pastoris, E. coli cannot perform higher eukaryotic post-translational modifications. As a result, proteins requiring these modifications often fold incorrectly when produced by E. coli. These disadvantages, which result from E. coli’s prokaryotic nature, limit the types of proteins that this system can express. Proteins that contain disulfide bonds or require post-translational modifications such as glycosylation, isomerization, or phosphorylation are not always properly expressed; they can be insoluble or improperly folded, which require additional solubilization and re-folding steps (Daly et al). Take for instance erythropoietin (EPO), a glycosylated protein. When EPO is expressed in E. coli, it is not glycosylated and becomes less resistant to unfolding compared to its natural, glycosylated form (Daly et al). Therefore, additional steps are needed in order to form a stable protein, which is time-consuming, costly, and has the tendency to produce low yields. <br />
<br />
A study done by Leuking et al. also supports the use of P. pastoris for protein expression. Vectors were created for both E. coli and P. pastoris using multiple cDNAs from a human fetal brain expression library. Out of the 29 DNA clones, all produced soluble proteins in P. pastoris, while E. coli was much less successful. In E. coli, only nine produced soluble proteins; 15 were detected as inclusion bodies; and five were not expressed at all. These differences are likely due to E. coli’s lack of eukaryotic abilities to perform post-translational modifications and properly fold proteins (Leuking et al).<br />
<br />
b. Powerful promoter systems<br />
<br />
P. pastoris’s wide range of promotors also aid in its use for foreign protein production. It has many strongly induced promotors, which allow sufficient expression of the genes of interest and therefore, a high concentration of proteins can be easily and inexpensively produced (Weidner et al). The alcohol oxidase genes, AOX1 and AOX2, are frequently used promotors because they are easily induced by methanol; because P. pastoris is methylotropic, meaning it can metabolize methanol as its carbon and energy source, adding methanol to the growth medium will induce these two promotors and then express the desired protein (Macauley-Patrick et al). High concentrations of protein can then be produced.<br />
A third advantage that P. pastoris has over E. coli is that proteins expressed by P. pastoris are easily purified (Weidner et al). Because P. pastoris secretes its recombinant protein in the growth medium and secretes low levels of endogeneous proteins, there is a better chance of obtaining high yields of uncontaminated proteins in comparison to E. coli (Weidner et al). In essence, there are fewer steps required to obtain the desired protein.<br />
<br />
<br />
Works Cited<br />
Daly, Rachel, and Milton T. W. Hearn. "Expression of Heterologous Proteins in Pichia pastoris: a Useful Experimental Tool in Protein Engineering and Production." PubMed.gov. PubMed, 26 Nov. 2004. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15565717>.<br />
Leuking, Angelika, Caterina Holz, Christine Gotthold, Hans Lehrach, and Dolores Cahill. “A System for Dual Protein Expression in Pichia pastoris and Escherichia coli.” PubMed.gov. PubMed, 15 Dec. 2003. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
Macauley-Patrick, Sue, Mariana L. Fazenda, Brian McNeil, and Linda M. "Heterologous Protein Production Using the Pichia pastoris Expression System." PubMed.gov. PubMed, 22 Mar. 2005. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15704221>.<br />
Weidner, Maria, Marcus Taupp, and Steven J. Hallam. "Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris." PubMed.gov. PubMed, 25 Feb. 2010. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/WhyPichiaTeam:Georgia State/WhyPichia2010-10-28T02:41:35Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
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<br />
[[Image:Whypichia.png|500px|right]]<br />
<br />
<br />
=='''Pichia Vs Saccharomyces'''== <br />
<br />
<br />
<br />
a. Greater Cell Concentrations<br />
The preference of Pichia pastoris for respiratory growth is a key characteristic which allows it to be cultured at high cell densities (500 OD600 U ml31)[1]. This trait lends an advantage to Pichia over Saccharomyces cervisiae: Resultant ethanol from S. cervisiae fermentation accumulates which hinders culture growth and therefore protein production. High levels of cell growth in fermenter cultures are vital to foreign protein production since concentration of the secreted product in "extracellular medium is...proportional to the concentration of cells in the culture". [2] <br />
<br />
<br />
=='''Easy and Inexpensive to Culture'''==<br />
<br />
<br />
The components of P. pastoris media (glycerol, methanol, salts, trace elements, and biotin) are relatively inexpensive and as such are well suited for large-scale heterologous protein production. The common protocol is to allow P. pastoris to accumulate biomass in a glycerol/carbon-source medium while foreign protein expression is repressed. Once glycerol stocks have been depleted, methanol is added to induce expression. Any undesirable protease activity can be easily averted by adjusting pH. Pichia's broad spectrum of optimal growth pH (3.0-7.0) provides ample wiggle room. A number of other techniques may be employed in minimizing proteolysis including lowering growth rate via methanol-level modulation, addition of protease inhibitors, addition of alternative protease substrates, and even simply lowering process temperature.<br />
<br />
<br />
== '''High production of foreign Protein while low levels of endogenous protein''' ==<br />
<br />
<br />
Extracellular expression of foreign proteins is favorable to intracellular production due to Pichia's extremely low levels of endogenous protein secretion, which makes it easier to isolate the protein of interest. Pichia vectors can also be cloned with foreign genes which align with a secretion signal (native to the protein, native to Pichia, or native to Saccharomyces), facilitating extracellular production. However, intracellular protein production is also high due to the efficiency--at carefully monitored methanol levels--of the AOX promoter. <br />
<br />
<br />
[1] Cregg 1999 review paper<br />
<br />
[2] Heterologous protein production using the Pichia pastoris expression system (2005). Sue Macaulet-Patric, mariana L. Fazenda, Brian McNeil, and Linda M. Harvey. Yeast (22): 249-270<br />
<br />
<br />
<br />
== '''Pichia pastoris versus Esherichia coli''' ==<br />
<br />
<br />
The bacterium E. coli has been frequently chosen as a host to express recombinant, heterologous proteins. Although many successful experiments have used E. coli as their expression system, P. pastoris has become an increasingly popular alternative due to the shortcomings of E. coli. P. pastoris serves as a better host when attempting to express foreign genes because this yeast can perform post-translational modifications and fold proteins properly, tightly regulate transcription via its wide range of promotors, and be purified easily.<br />
<br />
a. Post-translational modifications<br />
<br />
Unlike P. pastoris, E. coli cannot perform higher eukaryotic post-translational modifications. As a result, proteins requiring these modifications often fold incorrectly when produced by E. coli. These disadvantages, which result from E. coli’s prokaryotic nature, limit the types of proteins that this system can express. Proteins that contain disulfide bonds or require post-translational modifications such as glycosylation, isomerization, or phosphorylation are not always properly expressed; they can be insoluble or improperly folded, which require additional solubilization and re-folding steps (Daly et al). Take for instance erythropoietin (EPO), a glycosylated protein. When EPO is expressed in E. coli, it is not glycosylated and becomes less resistant to unfolding compared to its natural, glycosylated form (Daly et al). Therefore, additional steps are needed in order to form a stable protein, which is time-consuming, costly, and has the tendency to produce low yields. <br />
<br />
A study done by Leuking et al. also supports the use of P. pastoris for protein expression. Vectors were created for both E. coli and P. pastoris using multiple cDNAs from a human fetal brain expression library. Out of the 29 DNA clones, all produced soluble proteins in P. pastoris, while E. coli was much less successful. In E. coli, only nine produced soluble proteins; 15 were detected as inclusion bodies; and five were not expressed at all. These differences are likely due to E. coli’s lack of eukaryotic abilities to perform post-translational modifications and properly fold proteins (Leuking et al).<br />
<br />
b. Powerful promoter systems<br />
<br />
P. pastoris’s wide range of promotors also aid in its use for foreign protein production. It has many strongly induced promotors, which allow sufficient expression of the genes of interest and therefore, a high concentration of proteins can be easily and inexpensively produced (Weidner et al). The alcohol oxidase genes, AOX1 and AOX2, are frequently used promotors because they are easily induced by methanol; because P. pastoris is methylotropic, meaning it can metabolize methanol as its carbon and energy source, adding methanol to the growth medium will induce these two promotors and then express the desired protein (Macauley-Patrick et al). High concentrations of protein can then be produced.<br />
A third advantage that P. pastoris has over E. coli is that proteins expressed by P. pastoris are easily purified (Weidner et al). Because P. pastoris secretes its recombinant protein in the growth medium and secretes low levels of endogeneous proteins, there is a better chance of obtaining high yields of uncontaminated proteins in comparison to E. coli (Weidner et al). In essence, there are fewer steps required to obtain the desired protein.<br />
<br />
<br />
Works Cited<br />
Daly, Rachel, and Milton T. W. Hearn. "Expression of Heterologous Proteins in Pichia pastoris: a Useful Experimental Tool in Protein Engineering and Production." PubMed.gov. PubMed, 26 Nov. 2004. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15565717>.<br />
Leuking, Angelika, Caterina Holz, Christine Gotthold, Hans Lehrach, and Dolores Cahill. “A System for Dual Protein Expression in Pichia pastoris and Escherichia coli.” PubMed.gov. PubMed, 15 Dec. 2003. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
Macauley-Patrick, Sue, Mariana L. Fazenda, Brian McNeil, and Linda M. "Heterologous Protein Production Using the Pichia pastoris Expression System." PubMed.gov. PubMed, 22 Mar. 2005. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15704221>.<br />
Weidner, Maria, Marcus Taupp, and Steven J. Hallam. "Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris." PubMed.gov. PubMed, 25 Feb. 2010. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/WhyPichiaTeam:Georgia State/WhyPichia2010-10-28T02:39:05Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
[[Image:Whypichia.png|500px?right]]<br />
<br />
<br />
=='''Pichia Vs Saccharomyces'''== <br />
<br />
<br />
<br />
a. Greater Cell Concentrations<br />
The preference of Pichia pastoris for respiratory growth is a key characteristic which allows it to be cultured at high cell densities (500 OD600 U ml31)[1]. This trait lends an advantage to Pichia over Saccharomyces cervisiae: Resultant ethanol from S. cervisiae fermentation accumulates which hinders culture growth and therefore protein production. High levels of cell growth in fermenter cultures are vital to foreign protein production since concentration of the secreted product in "extracellular medium is...proportional to the concentration of cells in the culture". [2] <br />
<br />
<br />
=='''Easy and Inexpensive to Culture'''==<br />
<br />
<br />
The components of P. pastoris media (glycerol, methanol, salts, trace elements, and biotin) are relatively inexpensive and as such are well suited for large-scale heterologous protein production. The common protocol is to allow P. pastoris to accumulate biomass in a glycerol/carbon-source medium while foreign protein expression is repressed. Once glycerol stocks have been depleted, methanol is added to induce expression. Any undesirable protease activity can be easily averted by adjusting pH. Pichia's broad spectrum of optimal growth pH (3.0-7.0) provides ample wiggle room. A number of other techniques may be employed in minimizing proteolysis including lowering growth rate via methanol-level modulation, addition of protease inhibitors, addition of alternative protease substrates, and even simply lowering process temperature.<br />
<br />
<br />
== '''High production of foreign Protein while low levels of endogenous protein''' ==<br />
<br />
<br />
Extracellular expression of foreign proteins is favorable to intracellular production due to Pichia's extremely low levels of endogenous protein secretion, which makes it easier to isolate the protein of interest. Pichia vectors can also be cloned with foreign genes which align with a secretion signal (native to the protein, native to Pichia, or native to Saccharomyces), facilitating extracellular production. However, intracellular protein production is also high due to the efficiency--at carefully monitored methanol levels--of the AOX promoter. <br />
<br />
<br />
[1] Cregg 1999 review paper<br />
<br />
[2] Heterologous protein production using the Pichia pastoris expression system (2005). Sue Macaulet-Patric, mariana L. Fazenda, Brian McNeil, and Linda M. Harvey. Yeast (22): 249-270<br />
<br />
<br />
<br />
== '''Pichia pastoris versus Esherichia coli''' ==<br />
<br />
<br />
The bacterium E. coli has been frequently chosen as a host to express recombinant, heterologous proteins. Although many successful experiments have used E. coli as their expression system, P. pastoris has become an increasingly popular alternative due to the shortcomings of E. coli. P. pastoris serves as a better host when attempting to express foreign genes because this yeast can perform post-translational modifications and fold proteins properly, tightly regulate transcription via its wide range of promotors, and be purified easily.<br />
<br />
a. Post-translational modifications<br />
<br />
Unlike P. pastoris, E. coli cannot perform higher eukaryotic post-translational modifications. As a result, proteins requiring these modifications often fold incorrectly when produced by E. coli. These disadvantages, which result from E. coli’s prokaryotic nature, limit the types of proteins that this system can express. Proteins that contain disulfide bonds or require post-translational modifications such as glycosylation, isomerization, or phosphorylation are not always properly expressed; they can be insoluble or improperly folded, which require additional solubilization and re-folding steps (Daly et al). Take for instance erythropoietin (EPO), a glycosylated protein. When EPO is expressed in E. coli, it is not glycosylated and becomes less resistant to unfolding compared to its natural, glycosylated form (Daly et al). Therefore, additional steps are needed in order to form a stable protein, which is time-consuming, costly, and has the tendency to produce low yields. <br />
<br />
A study done by Leuking et al. also supports the use of P. pastoris for protein expression. Vectors were created for both E. coli and P. pastoris using multiple cDNAs from a human fetal brain expression library. Out of the 29 DNA clones, all produced soluble proteins in P. pastoris, while E. coli was much less successful. In E. coli, only nine produced soluble proteins; 15 were detected as inclusion bodies; and five were not expressed at all. These differences are likely due to E. coli’s lack of eukaryotic abilities to perform post-translational modifications and properly fold proteins (Leuking et al).<br />
<br />
b. Powerful promoter systems<br />
<br />
P. pastoris’s wide range of promotors also aid in its use for foreign protein production. It has many strongly induced promotors, which allow sufficient expression of the genes of interest and therefore, a high concentration of proteins can be easily and inexpensively produced (Weidner et al). The alcohol oxidase genes, AOX1 and AOX2, are frequently used promotors because they are easily induced by methanol; because P. pastoris is methylotropic, meaning it can metabolize methanol as its carbon and energy source, adding methanol to the growth medium will induce these two promotors and then express the desired protein (Macauley-Patrick et al). High concentrations of protein can then be produced.<br />
A third advantage that P. pastoris has over E. coli is that proteins expressed by P. pastoris are easily purified (Weidner et al). Because P. pastoris secretes its recombinant protein in the growth medium and secretes low levels of endogeneous proteins, there is a better chance of obtaining high yields of uncontaminated proteins in comparison to E. coli (Weidner et al). In essence, there are fewer steps required to obtain the desired protein.<br />
<br />
<br />
Works Cited<br />
Daly, Rachel, and Milton T. W. Hearn. "Expression of Heterologous Proteins in Pichia pastoris: a Useful Experimental Tool in Protein Engineering and Production." PubMed.gov. PubMed, 26 Nov. 2004. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15565717>.<br />
Leuking, Angelika, Caterina Holz, Christine Gotthold, Hans Lehrach, and Dolores Cahill. “A System for Dual Protein Expression in Pichia pastoris and Escherichia coli.” PubMed.gov. PubMed, 15 Dec. 2003. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
Macauley-Patrick, Sue, Mariana L. Fazenda, Brian McNeil, and Linda M. "Heterologous Protein Production Using the Pichia pastoris Expression System." PubMed.gov. PubMed, 22 Mar. 2005. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/15704221>.<br />
Weidner, Maria, Marcus Taupp, and Steven J. Hallam. "Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris." PubMed.gov. PubMed, 25 Feb. 2010. Web. 25 Oct. 2010. <http://www.ncbi.nlm.nih.gov/pubmed/20186119>.<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/File:Whypichia.pngFile:Whypichia.png2010-10-28T02:16:32Z<p>Djsexton5: </p>
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<div></div>Djsexton5http://2010.igem.org/Team:Georgia_State/PartsTeam:Georgia State/Parts2010-10-28T01:56:33Z<p>Djsexton5: /* Parts */</p>
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<div>{{Georgia_State/Header}}<br />
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===Pichia Tool Kit===<br />
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New for iGEM 2010 is the ''groupparts'' tag. <br />
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'''Welome to the Pichia Tool Box!!!''' Here you will find a selection of parts designed to facilitate your specific application of ''Pichia Pastoris''![[Image:toolbox.jpg|right]]<br />
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''Pichia pastoris'' has quickly gained attention as a powerful expression system in the last two decades. Researchers have successfully produced high yields of hundreds of heterologous protein from all orders of life. However, this incredible resource is yet to be tapped into by iGEM! We believe iGEM can play an enormous role in both the application and improvement of this expression system. In our tool box, we have standardized some of the most powerful, reliable and effective parts used today in ''P. pastoris''. From these parts, the ground work is laid for teams to get a head start to optimizing their particular application. We encourage you to experiment, explore and improve this system!<br />
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'''Part Design''': Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part. <br />
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[[Image:Toolbox Methods.png|center|700px]]<br />
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Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.<br />
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<groupparts>iGEM010 Georgia_State</groupparts></div>Djsexton5http://2010.igem.org/Team:Georgia_State/AboutthetoolboxTeam:Georgia State/Aboutthetoolbox2010-10-28T01:53:15Z<p>Djsexton5: </p>
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<div>{{Georgia_State/Header}}<br />
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'''How we made our parts'''<br />
<br />
Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part.<br />
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[[Image:Toolbox Methods.png|center|700px]]<br />
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[[Image:iGEMgel.jpg|center|400px]]<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/AboutthetoolboxTeam:Georgia State/Aboutthetoolbox2010-10-28T01:47:02Z<p>Djsexton5: </p>
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<div>{{Georgia_State/Header}}<br />
<br />
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'''How we made our parts'''<br />
<br />
Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part.<br />
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[[Image:Toolbox Methods.png|center|700px]]<br />
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[[Image:iGEMgel.jpg|300px.]]<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/File:IGEMgel.jpgFile:IGEMgel.jpg2010-10-28T01:28:25Z<p>Djsexton5: </p>
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<div></div>Djsexton5http://2010.igem.org/Team:Georgia_State/AboutthetoolboxTeam:Georgia State/Aboutthetoolbox2010-10-28T01:24:02Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
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'''How we made our parts'''<br />
<br />
Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/AboutthetoolboxTeam:Georgia State/Aboutthetoolbox2010-10-28T01:21:36Z<p>Djsexton5: New page: '''How we made our parts''' Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood ...</p>
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<div>'''How we made our parts'''<br />
<br />
Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part.</div>Djsexton5http://2010.igem.org/File:AOX1primer.pngFile:AOX1primer.png2010-10-28T01:02:33Z<p>Djsexton5: </p>
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<div></div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:41:29Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
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<div>{{Georgia_State/Header}}<br />
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==2010 Georgia State iGEM project==<br />
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'''Purpose #1: Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|300px]]<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|170px]] <br />
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[[Image:His4.png|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|170px]]<br />
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'''Purpose #2: Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|right|700px]]<br />
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{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:39:26Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
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<div>{{Georgia_State/Header}}<br />
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==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|300px]]<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|170px]] <br />
<br />
<br />
[[Image:His4.png|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|170px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:37:40Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
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==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|300px]]<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|right|170px]] <br />
<br />
<br />
[[Image:His4.png|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|170px]]<br />
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<br />
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<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:29:37Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
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<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|300px]]<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|right|170px]] <br />
<br />
[[Image:His4.png|left|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:26:24Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' <br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|right|170px]] <br />
[[Image:Pichia_tool_box_picture.jpg|right|300px]]<br />
[[Image:His4.png|left|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:22:38Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' <br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|left|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|right|170px]] [[Image:His4.png|left|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|left|100px]]<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:21:35Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' <br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|right|170px]] [[Image:FLD1.png|right|170px]] [[Image:HA antigen.png|right|170px]] [[Image:His4.png|right|170px]] [[Image:pGAP.png|Right|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
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[[Image:Pichia_tool_box_picture.jpg|left|100px]]<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:21:01Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
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<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' <br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
[[Image:AOX1.png|right|170px]] [[Image:FLD1.png|right|170px]] <br />
<br />
[[Image:HA antigen.png|right|170px]] [[Image:His4.png|right|170px]]<br />
<br />
[[Image:pGAP.png|Right|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|left|100px]]<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:20:11Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
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<div>{{Georgia_State/Header}}<br />
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<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' <br />
[[Image:AOX1.png|right|170px]] [[Image:FLD1.png|right|170px]] <br />
<br />
[[Image:HA antigen.png|right|170px]] [[Image:His4.png|right|170px]]<br />
<br />
[[Image:pGAP.png|Right|170px]] [[Image:pTEF.png|right|170px]]<br />
<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
[[Image:Pichia_tool_box_picture.jpg|left|100px]]<br />
<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:18:23Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|100px]]<br />
<br />
<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|170px]] [[Image:FLD1.png|170px]] [[Image:HA antigen.png|170px]]<br />
<br />
[[Image:His4.png|170px]] [[Image:pGAP.png|170px]] [[Image:pTEF.png|170px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:16:43Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|100px]]<br />
<br />
<br />
<br />
The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]] [[Image:FLD1.png|200px]] [[Image:HA antigen.png|200px]] [[Image:His4.png|200px]] [[Image:pGAP.png|200px]] [[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:15:45Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit''' [[Image:Pichia_tool_box_picture.jpg|right|100px]]<br />
<br />
<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]] [[Image:FLD1.png|200px]] [[Image:HA antigen.png|200px]] [[Image:His4.png|200px]] [[Image:pGAP.png|200px]] [[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:09:28Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]] [[Image:FLD1.png|200px]] [[Image:HA antigen.png|200px]] [[Image:His4.png|200px]] [[Image:pGAP.png|200px]] [[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.jpg|700px]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|100px]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:08:24Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]] [[Image:FLD1.png|200px]] [[Image:HA antigen.png|200px]] [[Image:His4.png|200px]] [[Image:pGAP.png|200px]] [[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
[[Image:vaccine.png|700px]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|100px]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/File:Vaccine.jpgFile:Vaccine.jpg2010-10-27T23:07:09Z<p>Djsexton5: </p>
<hr />
<div></div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:02:25Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]] [[Image:FLD1.png|200px]] [[Image:HA antigen.png|200px]] [[Image:His4.png|200px]] [[Image:pGAP.png|200px]] [[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T23:00:34Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|150px]] [[Image:FLD1.png|150px]] [[Image:HA antigen.png|150px]] [[Image:His4.png|150px]] [[Image:pGAP.png|150px]] [[Image:pTEF.png|150px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T22:58:55Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|100px]][[Image:FLD1.png|100px]][[Image:HA antigen.png|100px]][[Image:His4.png|100px]][[Image:pGAP.png|100px]][[Image:pTEF.png|100px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/File:Vaccine.pngFile:Vaccine.png2010-10-27T22:55:49Z<p>Djsexton5: </p>
<hr />
<div></div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T22:44:57Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]][[Image:FLD1.png|200px]][[Image:HA antigen.png|200px]][[Image:His4.png|200px]][[Image:pGAP.png|200px]][[Image:pTEF.png|200px]]<br />
<br />
<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/ProjectProject2010-10-27T22:43:49Z<p>Djsexton5: /* 2010 Georgia State iGEM project */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
<br />
<br />
<br />
==2010 Georgia State iGEM project==<br />
<br />
'''Tool Kit'''<br />
<br />
[[Image:Pichia_pastoris.jpg|right]] The primary purpose of the Georgia State 2010 team is to standardize parts that will facilitate future use of methylotrophic yeast "Pichia pastoris" as a novel chassis for iGEM purposes. We are titling this assembly of Pichia relevant parts as the “Pichia Tool Kit”, which is intended to be a constantly growing and improving resource. So far parts have included promoters, auxotrophic-mutant complementary proteins, selectivity parts and an antigen for intended production. Below is a diagram illustrating the parts that are currently in progress. Click on each image to go directly to the specific tool and get more information. <br />
<br />
<br />
[[Image:AOX1.png|200px]][[Image:FLD1.png|200px]][[Image:HA antigen.png|200px]][[Image:His4.png|200px]]<br />
<br />
'''Vaccine Production'''<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]<br />
<br />
[[Image:GSUteamphoto.JPG|400px]] [[Image:GSUAdvisors.jpg|300px]]<br />
<br />
The Georgia State University team is composed of stellar undergraduate students (up) and advised by brilliant people (down). We are interested in playing with organisms that haven't traditionally been used for bioengineering.<br />
<br />
<br />
{{Georgia_State/Footer}}</div>Djsexton5http://2010.igem.org/Team:Georgia_State/PartsTeam:Georgia State/Parts2010-09-22T04:49:33Z<p>Djsexton5: /* Parts */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
===Parts===<br />
<br />
<br />
<br />
New for iGEM 2010 is the ''groupparts'' tag. <br />
<br />
Welome to the Pichia Tool Box!!! Here you will find a selection of parts designed to facilitate your specific application of ''Pichia Pastoris''![[Image:toolbox.jpg|right]]<br />
<br />
<br />
''Pichia pastoris'' has quickly gained attention as a powerful expression system in the last two decades. Researchers have successfully produced high yields of hundreds of heterologous protein from all orders of life. However, this incredible resource is yet to be tapped into by iGEM! We believe iGEM can play an enormous role in both the application and improvement of this expression system. In our tool box, we have standardized some of the most powerful, reliable and effective parts used today in ''P. pastoris''. From these parts, the ground work is laid for teams to get a head start to optimizing their particular application. We encourage you to experiment, explore and improve this system!<br />
<br />
Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.<br />
<br />
<groupparts>iGEM010 Georgia_State</groupparts></div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-21T18:25:20Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]''Pichia pastoris'' is a methylotrophic yeast used as an alternative host for protein production in addition to ''Escherichia coli'' and ''Saccharomyces cerevisiae''. There are several reasons why ''P. Pastoris'' is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-21T18:22:51Z<p>Djsexton5: </p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]''Pichia pastoris'' is a methylotrophic yeast used as an alternative host for protein production in addition to ''Escherichia coli'' and ''Saccharomyces cerevisiae''. There are several reasons why ''P. Pastoris'' is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-21T18:18:58Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]''Pichia pastoris'' is a methylotrophic yeast used as an alternative host for protein production in addition to ''Escherichia coli'' and ''Saccharomyces cerevisiae''. There are several reasons why ''P. Pastoris'' is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, ''P.pastoris'' has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes ''P. pastoris'' would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in ''P. pastoris'' as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential ''P. pastoris'' has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/PartsTeam:Georgia State/Parts2010-09-21T18:17:10Z<p>Djsexton5: /* Parts */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
===Parts===<br />
<br />
<br />
<br />
New for iGEM 2010 is the ''groupparts'' tag. <br />
<br />
Welome to the Pichia Tool Box!!! Here you will find a selection of parts designed to facilitate your specific application of ''Pichia Pastoris''![[Image:toolbox.jpg|right]]<br />
Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.<br />
<br />
<groupparts>iGEM010 Georgia_State</groupparts></div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:31:49Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/Parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:22:54Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia_State/parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:22:12Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current toolbox here !! [[Team:Georgia State/parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:21:54Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current toolbox here !![[Team:Georgia State/parts]]<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:20:49Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current [[Team:Georgiastate/part]] !!<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:20:33Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current [[Team:Georgiastate/parts]] !!<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:20:14Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current [[Team:Georgia_state/parts]] !!<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/ProjectTeam:Georgia State/Project2010-09-07T00:17:54Z<p>Djsexton5: /* Pichia Pastoris as a host organism for iGEM */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
Over the past month, the team has worked diligently in preparing P. pastoris and Cyanobacteria competency, transforming DNA BioBrick parts, and optimizing growth conditions for future transformations. In addition, the group convened for the first annual GSU iGEM Boot Camp to work on team building, understanding the basic concepts, and organizing public relations.<br />
<br />
==Pichia Pastoris as a host organism for iGEM==<br />
[[Image:Pichia_pastoris.jpg|right]]Pichia pastoris is a methylotrophic yeast used as an alternative host for protein production in addition to Escherichia coli and Saccharomyces cerevisiae. There are several reasons why P. Pastoris is an ideal host organism. Its ability to perform eukaryotic post-translational modifications, high yields of recombinant protein, and its genetic similarity to Saccharomyces cerevisiae are very attractive traits(Cereghino and Cregg, 2000). <br />
<br />
<br />
Because of these reasons, P.pastoris has quickly gained popularity for recombinant protein production. The Georgia State 2010 team believes P. pastoris would be an excellent chassis for the iGEM competition. Our goal is to provide a tool box of parts necessary for the genetic manipulation of this organism. Parts will include a plasmid backbone, several parts providing alternative selectivity options and promoter systems. In addition, our tool box will be used to produce a flu virus antigen in P. pastoris as an example of how this system could be used for vaccine production. We hope our contributions will enable future users to maximize the use and further explore the incredible potential P. pastoris has to offer!<br />
<br />
Check out the current [[toolbox]] !!<br />
<br />
[[Image:Pichia_tool_box_picture.jpg|right]]</div>Djsexton5http://2010.igem.org/Team:Georgia_State/PartsTeam:Georgia State/Parts2010-09-07T00:15:08Z<p>Djsexton5: /* Parts */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
===Parts===<br />
<br />
<br />
<br />
New for iGEM 2010 is the ''groupparts'' tag. <br />
<br />
Welome to the Pichia Tool Box!!! Here you will find a selection of parts designed to facilitate your specific application of Pichia Pastoris![[Image:toolbox.jpg|right]]<br />
Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.<br />
<br />
<groupparts>iGEM010 Georgia_State</groupparts></div>Djsexton5http://2010.igem.org/Team:Georgia_State/PartsTeam:Georgia State/Parts2010-09-07T00:14:47Z<p>Djsexton5: /* Parts */</p>
<hr />
<div>{{Georgia_State/Header}}<br />
<br />
<br />
<br />
===Parts===<br />
<br />
<br />
<br />
New for iGEM 2010 is the ''groupparts'' tag. Welome to the Pichia Tool Box!!! Here you will find a selection of parts designed to facilitate your specific application of Pichia Pastoris![[Image:toolbox.jpg|right]]<br />
Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.<br />
<br />
<groupparts>iGEM010 Georgia_State</groupparts></div>Djsexton5http://2010.igem.org/File:Toolbox.jpgFile:Toolbox.jpg2010-09-07T00:13:54Z<p>Djsexton5: </p>
<hr />
<div></div>Djsexton5