Team:Northwestern/Project/Apoptosis

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<!--- The Mission, Experiments --->
<!--- The Mission, Experiments --->
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{|- style="font-size: 87%; background: #EFCDF8; color: #FFFFFF" cellpadding="1" cellspacing="1" bordercolor="#000000" width="75%" align="center"
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!align="center"|[[Team:Northwestern|<span style="color:#000000;">'''Home'''</span>]]
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!align="center"|[[Team:Northwestern|<font color="#2B3856">'''Home'''</font>]]
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!align="center"|[[Team:Northwestern/Brainstorm|<span style="color:#000000;">'''Brainstorm'''</span>]]
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!align="center"|[[Team:Northwestern/Brainstorm|<font color="#000000">'''Brainstorm'''</font>]]
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!align="center"|[[Team:Northwestern/Team|<span style="color:#000000;">'''Team'''</span>]]
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!align="center"|[[Team:Northwestern/Team|<font color="#000000">'''Team'''</font>]]
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!align="center"|[[Team:Northwestern/Acknowledgements|<span style="color:#000000;">'''Acknowledgements'''</span>]]
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!align="center"|[[Team:Northwestern/Acknowledgements|<font color="#000000">'''Acknowledgements'''</font>]]
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!align="center"|[[Team:Northwestern/Project|<span style="color:#000000;">'''Project'''</span>]]
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!align="center"|[[Team:Northwestern/Project|<font color="#000000">'''Project'''</font>]]
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!align="center"|[[Team:Northwestern/SideProject|<span style="color:#000000;">'''Side Project'''</span>]]
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!align="center"|[[Team:Northwestern/SideProject|<font color="#000000">'''Human Practices'''</font>]]
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!align="center"|[[Team:Northwestern/Parts|<span style="color:#000000;">'''Parts'''</span>]]
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!align="center"|[[Team:Northwestern/Parts|<font color="#000000">'''Parts'''</font>]]
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!align="center"|[[Team:Northwestern/Notebook|<span style="color:#000000;">'''Notebook'''</span>]]
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!align="center"|[[Team:Northwestern/Notebook|<font color="#000000">'''Notebook'''</font>]]
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!align="center"|[[Team:Northwestern/Calendar|<span style="color:#000000;">'''Calendar'''</span>]]
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!align="center"|[[Team:Northwestern/Calendar|<font color="#000000">'''Calendar'''</font>]]
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!align="center"|[[Team:Northwestern/Protocol|<span style="color:#000000;">'''Protocol'''</span>]]
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!align="center"|[[Team:Northwestern/Protocol|<font color="#000000">'''Protocol'''</font>]]
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!align="center"|[[Team:Northwestern/Safety|<span style="color:#000000;">'''Safety'''</span>]]
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!align="center"|[[Team:Northwestern/Safety|<font color="#000000">'''Safety'''</font>]]
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!align="center"|[[Team:Northwestern/Links|<span style="color:#000000;">'''Links'''</span>]]
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!align="center"|[[Team:Northwestern/Links|<font color="#000000">'''Links'''</font>]]
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!align="center"|[[Team:Northwestern/References|<span style="color:#000000;">'''References'''</span>]]
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!align="center"|[[Team:Northwestern/References|<font color="#000000">'''References'''</font>]]
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!align="center"|[[Team:Northwestern/Media|<span style="color:#000000;">'''Media'''</span>]]
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!align="center"|[[Team:Northwestern/Media|<font color="#000000">'''Media'''</font>]]
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!align="center"|[[Team:Northwestern/Contact|<span style="color:#000000;">'''Contact'''</span>]]
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!align="center"|[[Team:Northwestern/Contact|<font color="#000000">'''Contact'''</font>]]
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===T4 Holin===
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{| align="center" border="0" width="75%"
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|colspan="2"|
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===Endolycin===
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<table align="center">
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<td><a href="https://2010.igem.org/Team:Northwestern/Project/Modeling"><img width="100" class="icon" src="https://static.igem.org/mediawiki/2010/a/a7/NU_modeling.jpg"></a></td>
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<td><a href="https://2010.igem.org/Team:Northwestern/Project/Chassis"><img width="100" class="icon" src="https://static.igem.org/mediawiki/2010/a/a6/Nuchassis.jpg"></a></td>
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<td><a href="https://2010.igem.org/Team:Northwestern/Project/Lac"><img width="100" class="icon" src="https://static.igem.org/mediawiki/2010/1/18/Nulac.jpg"></a></td>
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<td><a href="https://2010.igem.org/Team:Northwestern/Project/Chitin Synthesis"><img width="100" class="icon" src="https://static.igem.org/mediawiki/2010/4/4e/Nuchitin.jpg"></a></td>
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<td><a href="https://2010.igem.org/Team:Northwestern/Project/Apoptosis"><img width="100" class="icon" src="https://static.igem.org/mediawiki/2010/c/cb/Nuapop.jpg"></a></td>
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</tr>
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<td align="center">Modeling</td>
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<td align="center">Chassis</td>
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<td align="center">Induction</td>
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<td align="center">Chitin</td>
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<td align="center">Apoptosis</td>
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===Rz Protein===
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===Live/Dead E.Coli Assay===
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=='''Apoptosis'''==
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Fluoresceinediacetate = stains living cells
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We used Part BBa_K124017, Bacteriophage Lysis Cassette, created by the Brown'08 iGEM team. The bacteriophage cassette includes the S105 protein (Holin), R protein (Endolysin), and Rz protein, all of which act in combination to lyse the cell. We attached the cassette to a promoter for it to be activated to produce proteins for cell lysis.
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Ethidium Bromide = stains dead cells
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Holin is a small membrane protein that produces holes in the membrane. This particular Holin protein contains the mutant S gene. Part BBa_K124014, Bacteriophage Holin Gene pS105, was also made by the Brown'08 iGEM team. The S105 gene has the initial 6 base pairs at the beginning of the wild type S gene sequence removed, which causes cell lysis to be induced at a faster rate than the wild type.
 +
 
 +
R protein is an endolysin that degrades the peptidoglycan cell wall, resulting in lysis of the cell by cleavage of the cell wall.
 +
 
 +
Rz protein, a periplasmic protein, carries out the final step of the cell lysis. The mechanism still has yet to be elucidated. However, it is clear that it is crucial to the function of the cassette.
 +
 
 +
When the Bacteriophage Lysis Cassette is activated, it will lyse the cell walls to release chitin that is produced via a slow mechanism.
 +
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Latest revision as of 18:33, 23 October 2010


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Modeling Chassis Induction Chitin Apoptosis

Apoptosis

We used Part BBa_K124017, Bacteriophage Lysis Cassette, created by the Brown'08 iGEM team. The bacteriophage cassette includes the S105 protein (Holin), R protein (Endolysin), and Rz protein, all of which act in combination to lyse the cell. We attached the cassette to a promoter for it to be activated to produce proteins for cell lysis.

Holin is a small membrane protein that produces holes in the membrane. This particular Holin protein contains the mutant S gene. Part BBa_K124014, Bacteriophage Holin Gene pS105, was also made by the Brown'08 iGEM team. The S105 gene has the initial 6 base pairs at the beginning of the wild type S gene sequence removed, which causes cell lysis to be induced at a faster rate than the wild type.

R protein is an endolysin that degrades the peptidoglycan cell wall, resulting in lysis of the cell by cleavage of the cell wall.

Rz protein, a periplasmic protein, carries out the final step of the cell lysis. The mechanism still has yet to be elucidated. However, it is clear that it is crucial to the function of the cassette.

When the Bacteriophage Lysis Cassette is activated, it will lyse the cell walls to release chitin that is produced via a slow mechanism.

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