Recombinational transfer of 100-kilobase genomic DNA to plasmid in Bacillus subtilis 168

K. Tsuge, M. Itaya

Research output: Contribution to journalArticle

28 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)5453-5458
Number of pages6
JournalJournal of Bacteriology
Volume183
Issue number18
DOIs
Publication statusPublished - 2001

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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