In Vivo Cloning of Arylsulfatase-Tyramine Oxidase Genes in rec Strains of Klebsiella aerogenes

Yoshikatsu Murooka, Masanori Sugiyama, Masahide Oka, Mitsuo Yamashita, Tokuya Harada

Research output: Contribution to journalArticle

Abstract

A general, genetic technique for in vivo cloning of bacterial genes is presented. We previously introduced the Mu phage into various genera of bacteria including Klebsiella aerogenes with RP4:: Mu. Using these strains carrying RP4:: Mu cts and thermo-inducible Mu prophage in the chromosome, we cloned in vivo the arylsulfatase (ats) and tyramine oxidase (tyn) genes by partial thermo-induction. The donor strains carrying the recombinant plasmids were conjugated with K. aerogenes rec strains, which were isolated as UV-sensitive mutants. The resultant recombinant plasmids, pATl and pAT2, were purified and used for the transformation of mutant strains deficient in the ats and tyn genes. The ats-tyn genes seemed to be transposed into the RP4::Mu cts plasmid together with other chromosomal DNA fragments. This in vivo cloning method is applicable to a wide variety of gram-negative bacteria.

Original languageEnglish
Pages (from-to)1807-1815
Number of pages9
JournalAgricultural and Biological Chemistry
Volume47
Issue number8
DOIs
Publication statusPublished - 1983
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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