Affiliation:
1. Mitsubishi Kasei Institute of Life Sciences, Machida-Shi, Tokyo 194-8511, Japan
Abstract
ABSTRACT
Transformation of
Bacillus subtilis
by a plasmid requires a circular multimeric form. In contrast, linearized plasmids can be circularized only when homologous sequences are present in the host genome. A recombinational transfer system was constructed with this intrinsic
B. subtilis
recombinational repair pathway. The vector, pGETS103, a derivative of the θ-type replicating plasmid pTB19 of thermophilic
Bacillus
, had the full length of
Escherichia coli
plasmid pBR322. A multimeric form of pGETS103 yielded tetracycline-resistant transformants of
B. subtilis
. In contrast, linearized pGETS103 gave tetracycline-resistant transformants only when the recipient strain had the pBR322 sequence in the genome. The efficiency and fidelity of the recombinational transfer of DNAs of up to 90 kb are demonstrated.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
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