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From 2012.igem.org

Achievements and Failures

Successfully cloned and submitted 32 biobricks into the registry Obtained wbbC gene from BL21 genome Created GlycoBase; a database containing a list of >100 glycosyltransferase enzymes from E.coli strains to demonstrate the vast linkages we could achieve only with E.coli enzymes Created GlycoWeb, an interface for an online user to access Glycobase with a polysaccharide in mind and the database searches and finds the enzymes you’d use to make it. Created GlycoApp; an application for a handheld device to access the Glycobase from wherever the consumer is when they realise they need a polysaccharide ordering! Considered human practice approaches and discussed with several experts the human practice considerations, hosted a human practice panel and discussed with the wider community at Café Scientifique. Discussed our project with various businesses including Shell, Ginsters etc. and we received letters of support from DSTL, ISCA Biochemical Ltd, Avon. Demonstrated that….. Successfully expressed this polysaccharide….. (to be cont….) Made promoter-RBS-gene-terminator constructs for each gene A correct mass analysis of the product is essential, but cannot alone confirm the correct product. The possibility that an incorrect product has been made through incorrect synthesis or post-synthetic regiospecific or enantiospecific rearrangement by chemical or enzymatic means must be examined and ruled out before the method can be considered reliable. 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. Only managed to obtain 7 of 9 fragments for Gibson assembly through PCR. Didn’t manage to construct operons through BioBrick assembly.