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From 2012.igem.org

Achievements and Failures

Successfully cloned and submitted 14 biobricks into the registry Successfully cloned over 32 more Biobricks waiting to be transferred into pSB1C3 plasmids Using PCR we obtained wbbC gene from BL21 genome Created GlycoBase; a database containing a list of over 100 different glycosyltransferase enzymes mainly from E.coli strains demonstrating the vast linkages we could achieve with just E.coli enzymes Created GlycoWeb, an interface for an online user to access Glycobase. Glycoweb has the ability to tell you which enzymes are needed to create your bespoke polysaccharide. Created GlycoApp; an application for a handheld device to access the Glycobase giving the user distance access and enhanced ordering capabilities The project evolved with human practices for the consideration of the ethical, societal, environmental and business aspects of our project. We held a human practice panel, Café Scientifique talk, met with various company members from different business sectors, led an A level master class and worked with several work experience students. The discussions with various businesses had a very positive response and we received several letters of support. We created devices to test our proteins, test our natural polysaccharides and to help construct our 3 gene inducible plasmid. These included BBa_K094120_BBa_B0034_wclY_BBaB0014, BBa_J13002_wbnk_BBaB0014, BBa_J23119_BBa_B0034_ wbnJ_BBaB0014 BBa_J13002_ompA_BBa_J322921_BBa_B0034 and BBa_K20600_ompA_BBa_J322921_BBa_B0034. We managed to obtain 7 of 9 fragments for Gibson assembly through PCR. The different annealing temperatures of the primers made this hard but showed how important the preparation for Gibson has to be In touching distance of completing our operons. Only time stopped this happening. We were one construct away from being able to complete our 3 gene inducible plasmid. Synthesis of a previously well characterised small polysaccharide, which can be physically and also biochemically characterised, to show that the product will act in biological systems as expected. Once a fully characterised polysaccharide is produced, we would be in a much stronger position to offer novel products and to start to address the exciting possibilities of our method. We were not able to fully test our constructs and properly characterise our biobricks.