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Team:Panama/Project
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The purpose behind our project is to take a gene that produces the Rhamnosyltransferase 1 enzyme in a pathogenic bactria known as ''Pseudomona aeruginosa'' and insert it into an efficient and not so pathogenic bacteria that is ''Escherichia coli''. This will help create a rhamnolipid that can aid in the more efficent degradation of hydrocarbons. | The purpose behind our project is to take a gene that produces the Rhamnosyltransferase 1 enzyme in a pathogenic bactria known as ''Pseudomona aeruginosa'' and insert it into an efficient and not so pathogenic bacteria that is ''Escherichia coli''. This will help create a rhamnolipid that can aid in the more efficent degradation of hydrocarbons. | ||
| - | We'll start out by selecting the components of our gene which will be inserted into the ''E. coli'' for expresion. We have chosen a promoter, a RBS, a reporter and a terminator. All of this from the plates that our friends at iGEM have conviniently sent all of us. These plasmids will then be used to obtain transformed bacteria that have those parts in them | + | We'll start out by selecting the components of our gene which will be inserted into the ''E. coli'' for expresion. We have chosen a promoter, a RBS, a reporter and a terminator. All of this from the plates that our friends at iGEM have conviniently sent all of us. These plasmids will then be used to obtain transformed bacteria that have those parts in them. Now, we can't use the entire plasmid for our construction. We have to extract that gene for the promoter, RBS, reporter and terminator from the plasmid to be able to use them, so we'll perform minipreparations to be able to achieve that. |
In the meantime, we have to obtain our gene from the ''P. aeruginosa''. This bacteria is donated to us by Marcelino Gutierrez, Ph.D, who is a researcher from the chemistry department at INDICASAT-AIP. They were given to us in liquid media ready for extraction of genomic DNA. Our DNA of interest wil be yielded by performing a PCR. | In the meantime, we have to obtain our gene from the ''P. aeruginosa''. This bacteria is donated to us by Marcelino Gutierrez, Ph.D, who is a researcher from the chemistry department at INDICASAT-AIP. They were given to us in liquid media ready for extraction of genomic DNA. Our DNA of interest wil be yielded by performing a PCR. | ||
Revision as of 15:08, 25 October 2010
Republic of Panama
Situated on the isthmus connecting North and South America, it
is bordered by Costa Rica to the northwest, Colombia to the southeast,
the Caribbean Sea to the north and the Pacific Ocean to the south.
Republic of Panama
Situated on the isthmus connecting North and South America, it is bordered by Costa Rica to the northwest, Colombia to the southeast, the Caribbean Sea to the north and the Pacific Ocean to the south.
iGEM PANAMA
In this picture: Carolina, Yisett, Claudio, Nicole, Lorena,
Grimaldo, Natasha, Laura, Ernesto, Dra. Carmenza, Dr. Rao, Ezequiel
iGEM PANAMA
In this picture: Carolina, Yisett, Claudio, Nicole, Lorena, Grimaldo, Natasha, Laura, Ernesto, Dra. Carmenza, Dr. Rao, Ezequiel
iGEM PANAMA
In this picture: Yisett, Silke, Yaraví, Carlos, Dr. Patrick
Nee.
iGEM PANAMA
In this picture: Yisett, Silke, Yaraví, Carlos, Dr. Patrick Nee.
iGEM PANAMA
In this picture: Leyda, Zeuz, Laura
iGEM PANAMA
In this picture: Leyda, Zeuz, Laura
iGEM PANAMA
Labs.
iGEM PANAMA
Labs.
Contents |
Overall project
Standardization of the Rhamnosiltransferase 1 gene complex (rhlAB) into a Biobrick-friendly for rhamnolipid production in E. coli.
There is considerable interest among bio-industries in bioremediation products such as Rhamnolipids. Rhamnolipids as biosurfactants are important in the remediation of oil spill areas. The cleanup of the Exxon Valdez oil spill with rhamnolipids as biosurfactants was too expensive and complicated, therefore impractical for large-scale bioremediation. However, advances in genetic engineering and synthetic biology offer a viable solution to oil spill pollution clean up. In this project we use genetic engineering as a tool to integrate genetic parts by the biobrick assembly standard protocol of iGEM to develop a biobrick-friendly for rhamnosiltransferase 1 complex (rhlAB) gene expression in Escherichia coli for standardized rhamnolipid production. Our biobrick integrates a promoter, a RBS (ribosomal binding site), our gene sequence of rh1AB isolated from Pseudomonas aeruginosa, a GFP reporter and a terminator. All the parts fit into a plasmid backbone that can be transformed into E. coli strains, which can then produce rhamnolipids.
Project Details
The Experiments
The purpose behind our project is to take a gene that produces the Rhamnosyltransferase 1 enzyme in a pathogenic bactria known as Pseudomona aeruginosa and insert it into an efficient and not so pathogenic bacteria that is Escherichia coli. This will help create a rhamnolipid that can aid in the more efficent degradation of hydrocarbons.
We'll start out by selecting the components of our gene which will be inserted into the E. coli for expresion. We have chosen a promoter, a RBS, a reporter and a terminator. All of this from the plates that our friends at iGEM have conviniently sent all of us. These plasmids will then be used to obtain transformed bacteria that have those parts in them. Now, we can't use the entire plasmid for our construction. We have to extract that gene for the promoter, RBS, reporter and terminator from the plasmid to be able to use them, so we'll perform minipreparations to be able to achieve that.
In the meantime, we have to obtain our gene from the P. aeruginosa. This bacteria is donated to us by Marcelino Gutierrez, Ph.D, who is a researcher from the chemistry department at INDICASAT-AIP. They were given to us in liquid media ready for extraction of genomic DNA. Our DNA of interest wil be yielded by performing a PCR.
Once we have this, we have to mutate the Rhamnosyltransferase 1 gene to eliminate the restriction site of Pst1 that clashes with the iGEM disposition that indicates that our gene cannot be cut in any way by the restriction enzymes. This mutation will allow us to have the same gene without the restriction sites within our construct permitted.
In the end, we'll be able to ligate all the parts together to have one general section that contains the Promoter+RBS+ OUR GENE +Reporter+Terminator
