Team:Edinburgh UG/Improve

Edinburgh iGEM 2018

Improve

BBa_K2725001

We improved the characterisation of a previous part: BBa_K771303, which contains the coding sequence of E. coli FabI. FabI is involved in type 2 fatty acid synthesis and has previously been characterised as a biosensor for triclosan, a common biocide. Overexpression of FabI has been shown to give resistance to triclosan, so we characterised the use of FabI as an alternative selection gene. Usage of triclosan instead of antibiotics is considerably cheaper, and safer for the environment.

BBa_K914009 & BBa_K914018

We improved the characterisation of the previous parts: BBa_K914009 & BBa_K914018, which contain a kanamycin resistance gene with one and two serine -> amber codon mutations, respectively. Previously, their kanamycin resistance with and without a suppressor tRNA were characterised in E. coli MG1655. We further characterised their ability to confer resistance in E. coli TOP10 and made further parts with a larger number of amber mutations.

BBa_K2725012 & BBa_K2725013

We improved the previous parts: BBa_K914009 & BBa_K914018, which contain a kanamycin resistance gene with one and two serine -> amber codon mutations, respectively. By increasing the number of serine -> amber codon mutations to 5 in BBa_K2725012 and to 10 in BBa_K2725013, they confer a decreased kanamycin resistance in the absence of a suppressor tRNA, reducing the leakiness of the gene and thereby increasing its safety.

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