Molecular cloning of the gene (poxB) encoding the pyruvate oxidase of Escherichia coli, a lipid-activated enzyme

C. Grabau, John E Cronan

Research output: Contribution to journalArticlepeer-review

Abstract

The pyruvate oxidase structural gene (poxB) of Escherichia coli was cloned into derivatives of plasmid pBR322. The gene was first cloned into a cosmid vector by selection for the tetracycline resistance determinant of a closely linked Tn10 insertion (no direct selection for the gene was available). Subsequent subcloning resulted in localization of the gene to a 3.1-kilobase-pair DNA segment. Two of the smaller poxB plasmids were shown to cause the overproduction of oxidase activity (by six- to eightfold), and one of these plasmids was shown to encode a protein having the size and antigenic determinants of pyruvate oxidase. Introduction of poxB plasmids into strains (aceEF) lacking pyruvate dehydrogenase activity relieved the aerobic growth requirement of the strains for exogenous acetate.

Original languageEnglish (US)
Pages (from-to)1088-1092
Number of pages5
JournalJournal of Bacteriology
Volume160
Issue number3
StatePublished - 1984

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

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