In fulfillment of the requirements for the Gold Medal, the 2009 Utah State iGEM Team did the following:
Bronze Medal Requirements
- Completed the registration requirements and Project Summary form.
- Prepared and will present a poster and talk at the 2009 Jamboree.
- Entered all necessary information detailing 62 BioBricks into the Registry of Standard Parts.
- Designed parts in conformity with accepted BioBrick standards.
- DNA for 49 BioBricks entered in the Registry were sent in to iGEM headquarters. The majority of these parts were confirmed with DNA sequencing.
Silver Medal Requirements
- Demonstrated that several submitted BioBricks work as expected.
- The composite lac promoter/RBS/GFP/terminator (BBa_K208045). This composite was demonstrated by the presence of GFP visualized on a UV transilluminator. The GFP protein was also visualized using SDS polyacrylamide gel electrophoresis. See pictures on Wiki.
- New GFP reporter (BBa_K208000). This reporter, with an excitation/emission of 395/509, was shown to be functional using the novel composite construct BBa_K2208045 (lac promoter/RBS/GFP/terminator). Picture on Wiki under part BBa_K2208045.
- The composite lac promoter/RBS (BBa_K208010). This was demonstrated using the composite construct BBa_K2208045. Picture on Wiki under part BBa_K2208045. This composite part is extremely useful because it alleviates the need to work with extremely small ribosomal BioBrick components.
- Lac promoter/RBS/Gene III/Phasin/terminator (BBa_K208038). This was demonstrated through the detection of phasin proteins isolated from supernatant samples using SDS polyacrylamide gel electrophoresis. Picture on Wiki.
- Phasin gene (BBa_K208001). A protein of the correct size was detected in SDS polyacrylamide gel electrophoresis from cells having this construct that was not detected in the control samples.
- Gene III secretion tag (BBa_K208002). The phasin protein from part BBa_K208038 was detected outside the cell, thus demonstrating the functionality of this Gene III secretion tag.
- Characterized the GFP reporter (BBa_K208000) (1) showing GFP expression over time and (2) showing sensitivity of IPTG induction.
Gold Medal Requirements
- Improved upon the existing set of GFP BioBrick parts by constructing a GFP with a larger Stokes shift (395/509nm), making for easier downsteam analysis. The bright fluorescence can very easily be seen on a standard UV transilluminator. As GFP is one of the most commonly used reporters, the introduction of this new and improved BioBrick part should be of great benefit to the iGEM community.
- Our initial plans for our 2009 team was in part to continue the 2008 Hawaii team’s project, “Cyanobacteria Toolkit,” by making the pRL1383a vector an effective broad-host vector. When this vector proved ineffective, we made efforts to troubleshoot. We had much correspondence with the advisors of the Hawaii team, including a conference call mid-summer. Since most of their parts were not included in the 2009 iGEM distribution, we had them send many of the parts they used. After numerous approaches and attempts to convert pRL1383a into BioBrick format, we finally decided that the vector was either mischaracterized or altered in some unknown way.
- As thoroughly explained in our Wiki, one of the primary goals of our project has been to expand the BioBrick world to organisms other than E. coli. We have documented the many benefits that would come from such an achievement. Though the pRL1383a vector efforts were unsuccessful, we did successfully express the vector pCPP33 in E. coli, Pseudomonas putida, Synechocystis pcc 6803, and Rhodobacter sphaeroides. This vector now has only to be put in BioBrick format.
- In reference to our ethics section (http://2009.igem.org/Team:Utah_State/ETHICS), our team has taken specific measures to follow our suggested proposals. In the education of our team, we discussed the potential benefits of a standard secretion system but also discussed the potential of our designed secretion pathways to be used in a malevolent manner. As a team, we acknowledge the importance of high moral accountability and commitment to safety and security. Additionally, in an effort to foster the sharing of information in our community, upon completion of the jamboree we will submit an article to be released in our school newspaper and in the College of Engineering website. This winter we are also hosting a lecture by Drew Endy addressing synthetic biology which will be open to the public. We hope to act as ambassadors to foster support and excitement in our own community.