Conjugative plasmid transfer from Escherichia coli to Clostridium acetobutylicum

Abstract
SUMMARY: The conjugation mechanism of IncP plasmids may be employed to mobilize small non-conjugative plasmids from Escherichia coli to a wide range of different organisms. This strategy has been adapted for use with the Grampositive anaerobe, Clostridium acetobutylicum NCIB 8052. Several shuttle vectors containing replicons from pAMß1 (Enterococcus faecalis), pCB101 (Clostridium butyricum) or pWV01 (Streptococcus cremoris), together with the cis-acting oriT region of RK2, have been constructed, and transferred to and established in this organism. One of the vectors apparently contains a hot-spot for insertion of IS1. Conjugative mobilization of plasmids from E. coli will provide a useful alternative to electroporation for effecting gene transfer to this industrially important anaerobe.