Electrotransformation of clostridium pasteurianum

摘要

A method for high-efficiency genetic transformation of the anaerobic bacterium Clostridium pasteurianum is provided. Clostridium pasteurianum is industrially important due to its selective and high level production of the biofuel and biochemical n-butanol, and its ability to grow on a wide variety of inexpensive substrates. C. pasteurianum can use as a sole source of carbon and energy is glycerine, which is a by-product of biodiesel processing. The industrial exploitation of Clostridium pasteurianum has previously been impeded by the lack of genetic engineering tools for this bacterium. The method provided enables high-efficiency transformation of C. pasteurianum via a series of treatments and electroporation conditions which successfully negotiate the resistant cell wall of C. pasteurianum, and includes a means for protecting newly introduced DNA from degradation by a restriction enzyme within C. pasteurianum as well as selection markers and vector components.