| 摘要 |
A method for producing a deletion in a bacterial genome, wherein the bacteria comprise a CRISPR/Cas system, comprises transforming the bacteria with one or more Deletion Vectors encoding first and second crRNAs targeting first and second PAM/Protospacers within the genome of the bacteria, resulting in dual cleavage of the genome with rejoining of the ends to produce a deletion of the region between the first and second PAM/Protospacers. A method is also provided for removing an endogenous plasmid within a bacteria, wherein the bacteria are transformed with one or more Deletion Vectors encoding two or more crRNAs targeting two or more PAM/Protospacers within the plasmid, resulting in the production of linearised fragments which subject to degradation by endogenous cell mechanisms. Preferably the bacteria are from the genus Clostridium, and possess a type I CRISPR/Cas system. |
