As this year's winners of the SYNtheSYS competition on synthetic and systems biology at Heidelberg University we will spend the next six weeks in a lab at BioQuant to realise our project. We - that is Katharina, Jan, Nikos, Johyo and Silvan - aim to develop an orthogonal T7 replication system in E.coli as a tool in directed evolution. This means, that we will spend hours at a time mixing microlite
rs of clear liquids, heating them, cooling them and shaking them. And to give us an excuse to check facebook whilst in the lab ;)
In a bit more detail:
Directed evolution has become a widely used tool to engineer improved enzymes. Most approaches are based on generating a gene library, e. by error-prone PCR. These libraries are then usually expanded further by recombination and other techniques. After transforming bacterial cultures with these genes, mutants with the desired improvement are chosen by means of screening or natural selection. Our goal is to establish a system that allows in vivo mutagenesis of specific target genes. In order to achieve this and in the spirit of synthetic biology we intend to construct standardized biological parts, biobricks of the essential components of the bacteriophage T7 replication apparatus including error-prone mutant of the T7 DNA polymerase and the T7 replication origin. Ideally, the T7 polymerase will only replicate the plasmid with the T7 origin and hence the mutagenesis induction will have a localized effect (high specificity). For further information check our team page on the SYNtheSYS website.
