Team:GeorgiaTech/Modeling

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Collaboration

Georgia State University

In a meeting with Georgia State University’s IGEM team, they informed us of their project to express the gene Mambalgin-1 in E.coli. Mambalgin is a powerful and non-addictive analgeasic isolated from black mamba venom; given it's eukaryotic origins, GSU has gone through the hoops to create a single functional E.coli expression vector for Mambalgin. In this year's project they aimed to both optimize and characterize the production of Mambalgin in E.coli. It was evident that they planned to spend much of the summer creating new varied strength expression vectors for Mambalgin. Having already seen promising results from our new insertion primers, we informed them that, using our revolutionary RBS and Promoter Primer PCR, we could add a ribosome binding site and a promoter to the Mambalgin gene both cheaper and faster. Their interest in our proposal thus led to the creation of several promoter and RBS insertion primers specifically for eukaryotic Yeast.

  • Forward Primer sequence(Adds Biobrick Prefix)

  • Reverse Primer sequence(Adds Biobrick Suffix)

  • PCR Product using the above two primers with the Mambalgin-1 template DNA

Together the primers amplified a 225 bp long segment of DNA as predicted. This PCR product was subsequently ligated into a backbone and transformed into E.coli; it was later confirmed successful by sequencing. Next, the ribosome binding site was inserted using RBS Primer PCR method that developed here at Georgia Tech. Since the start codon for in this case was 'CTG', a new RBS primer was designed with the same structure and components ( prefix, RBS, scar and start codon) as universal primers, but with ‘CTG’ as the ending base pairs. Success of this PCR ultimately confirmed the adaptability of the RBS insertion method.

  • RBS insertion primer sequence

  • New PCR product using the RBS insertion primer and the Reverse Primer from above (only prefix end shown)

Lambert Highschool

During the summer the Georgia Tech iGEM team aided in critiquing the Lambert High School iGEM team’s presentation for the manufacture of chitosan, which helps preserve fruit and vegetables from spoiling rapidly. On two different occasions, our research team oversaw the Lambert iGEM team’s presentation, helping them improve their presentation format, style, and slide orientation.

Our Team's Wiki Czar also contributed to the design, formatting, and HTML coding of Lambert High School's Wiki. Just three days before the iGEM 2014 High School Wiki freeze, Lambert High School's wiki was difficult navigate and with formatting and a color palette liable to give the reader a headache if stared at for too long. All of this was alleviated over the next 3 days as our Wiki Czar and the Lambert Team coded together to create a wiki that really captured the value of their work and that they could proudly display at their own iGEM Jamboree.

Click Here to See Lambert High School's 2014 iGEM Wiki

University of Virginia