ClosTron Mutagenesis


Directed mutagenesis is an important approach in molecular microbiology, but has historically proven difficult in many bacteria, including clostridial species. This led to the development of the ClosTron system, which allowed the directed construction of stable mutants in Clostridium species using the Ll.LtrB intron of Lactococcus lactis. The intron is directed to insert into the desired DNA target by altering the sequence of the intron RNA-encoding ClosTron plasmid.

The procedure for ClosTron mutagenesis has been optimised to exploit key characteristics of group II introns, facilitating mutant generation in Clostridia: