Genetic transformation ofClostridium botulinumhall a by electroporation

✍ Scribed by Yongtai Zhou; Eric A. Johnson

• Book ID: 104634821
• Publisher: Springer Netherlands
• Year: 1993
• Tongue: English
• Weight: 402 KB
• Volume: 15
• Category: Article
• ISSN: 0141-5492
• DOI: 10.1007/bf00133010

✦ Synopsis

A method was developed for the introduction of plasmids into Clostridium botulinurn by electroporation. A 4.4 kb plasmid vector, pGK12, which contains genes for resistance to erythromycin (Em') and chloramphenicol (Cmr) was electroporated into C. botulinurn type A (Hall A). The highest transformation efficiency was obtained using midlog phase cells, 10% PEG 8000 as the electroporation solution, and 2.5 kV field strength. The transformation efficiency was highest (-103 transformants&g of DNA) when 1 pg of plasmid DNA and 4 X 108 CFU/ml of recipient cells were used. Plasmid DNA recovered from the transformants was indistinguishable from that introduced on the basis of restriction enzyme digestion and agarose gel electrophoresis.