The plasmid being used is pBS1 and it has 2 segments of DNA on it which are derived from the B.subtilis amylase gene. There is a chloramphenicol resistance marker inserted between the amyfront and amyback arms. I know that if a single crossover occurs the entire plasmid is integrated and a copy of the amylase gene will remain intact, but not what would happen with a single crossover if the plasmid was linearised (cut). I need to know what will happen when this gap gets incorporated into the Bacillus chromosome and what happens to the bacterial cells as a result.
The linearized plasmid would cross over into the chromosome, producing a linearized chromosome with plasmid sequences at the ends. Cells would die unless they underwent non-homologous end-joining. If you had a second crossover within the plasmid, this would be a 'gene replacement' kind of effect.
The linearized plasmid would cross over into the chromosome, producing a linearized chromosome with plasmid sequences at the ends. Cells would die unless they underwent non-homologous end-joining. If you had a second crossover within the plasmid, this would be a 'gene replacement' kind of effect.
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