PLASMID TRANSFER INTO THE HOMOACETOGEN ACETOBACTERIUM-WOODII BY ELECTROPORATION AND CONJUGATION

Shuttle vectors (pMS3 and pMS4) which replicated in Escherichia coli a nd in gram-positive Acetobacterium woodii were constructed by ligating the replication origin of plasmid pAM beta 1 with the E. coli cloning vector pUC19 and the tetM gene of streptococcal transposon Tn916. Ele ctrotransformation of A. woodii was achieved at frequencies of 4.5 x 1 0(3) transformants per mu g of plasmid DNA. For conjugal plasmid trans fer, the mobilizable shuttle vector pKV12 was constructed by cloning t he tetM gene into pAT187. Mating of E. coli containing pKV12 with A. w oodii resulted in transfer frequencies of 3 x 10(-6) to 7 X 10(-6) per donor or recipient.