Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis

Lam Son Phan Tran, Lóránd Szabó, László Orosz, Tibor Sík, András Holczinger

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A versatile system consisting of an integrational vector and a bacitracin (Bt)-producing β-galactosidase (β-Gal)-negative (Lac-) Bacillus licheniformis TLH strain was constructed to quantify promoter activity and to study gene regulation in a single-copy set-up. The vector pTLH utilizes the promoterless Escherichia coli lacZ gene derived from pQF52 and contains the pBR322 origin of replication and a kanamycin-resistance gene for selection in both B. licheniformis and E. coli. The vector also contains an inner part of the first gene of the Bt synthetase (bts) operon which enables its integration into the bts of B. licheniformis by Campbell-type recombination. This recombination event can be easily tested on a Micrococcus flavus lawn where loss of Bt production, i.e. no clearing zone on the lawn, is indicative of the proper integration. The Lac- B. licheniformis TLH strain was developed by elimination of the natural β-Gal activity of B. licheniformis strain ATCC 10716 UM12 using NTG mutagenesis.

Original languageEnglish
Pages (from-to)2573-2578
Number of pages6
JournalMicrobiology
Volume144
Issue number9
DOIs
Publication statusPublished - Sep 1998

Keywords

  • Bacitracin synthetase
  • Campbell-type recombination
  • Gene expression
  • NTG mutagenesis
  • lacZ fusions

ASJC Scopus subject areas

  • Microbiology

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