Part:BBa_K374017:Experience

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Applications of BBa_K374017

A plasmid containing the arabinose inducible promoter pBAD was used to test the anti-repressor AntT. AntT was cloned into the plasmid downstream of the pBAD promoter and then the plasmid was transformed into the Salmonella enterica serovar Typhumurium strain which has the Gifsy2 phage incorporated in the genome.

The immunity region of the Gifsy2 consists of a pRM promoter with the repressor GtgR located downstream and a pR promoter which is regulated by GtgR. When the pR promoter is repressed by the GtgR repressor the lysogenic life cycle of the Gifsy2 phage is maintained.

After the transformation the cells were grown overnight. Then the pBAD promoter was induced by arabinose and as expected the cells lysed indicating that the phage had changed from lysogenic to lytic cycle.

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