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

Lam Son Phan Tran, Lóránd Szabó, L. 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
Publication statusPublished - Sep 1998

Fingerprint

Gene Expression
Bacitracin
Genetic Recombination
Galactosidases
Kanamycin Resistance
Escherichia coli
Genes
Micrococcus
Replication Origin
Lac Operon
Operon
Ligases
Mutagenesis
Bacillus licheniformis

Keywords

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

ASJC Scopus subject areas

  • Microbiology

Cite this

Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis. / Tran, Lam Son Phan; Szabó, Lóránd; Orosz, L.; Sík, Tibor; Holczinger, András.

In: Microbiology, Vol. 144, No. 9, 09.1998, p. 2573-2578.

Research output: Contribution to journalArticle

Tran, LSP, Szabó, L, Orosz, L, Sík, T & Holczinger, A 1998, 'Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis', Microbiology, vol. 144, no. 9, pp. 2573-2578.
Tran, Lam Son Phan ; Szabó, Lóránd ; Orosz, L. ; Sík, Tibor ; Holczinger, András. / Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis. In: Microbiology. 1998 ; Vol. 144, No. 9. pp. 2573-2578.
@article{785ead18b08649bf964270d900cb38a2,
title = "Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis",
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.",
keywords = "Bacitracin synthetase, Campbell-type recombination, Gene expression, lacZ fusions, NTG mutagenesis",
author = "Tran, {Lam Son Phan} and L{\'o}r{\'a}nd Szab{\'o} and L. Orosz and Tibor S{\'i}k and Andr{\'a}s Holczinger",
year = "1998",
month = "9",
language = "English",
volume = "144",
pages = "2573--2578",
journal = "Microbiology",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "9",

}

TY - JOUR

T1 - Construction of a single-copy integration vector and its use to study gene expression in Bacillus licheniformis

AU - Tran, Lam Son Phan

AU - Szabó, Lóránd

AU - Orosz, L.

AU - Sík, Tibor

AU - Holczinger, András

PY - 1998/9

Y1 - 1998/9

N2 - 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.

AB - 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.

KW - Bacitracin synthetase

KW - Campbell-type recombination

KW - Gene expression

KW - lacZ fusions

KW - NTG mutagenesis

UR - http://www.scopus.com/inward/record.url?scp=0031669914&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031669914&partnerID=8YFLogxK

M3 - Article

VL - 144

SP - 2573

EP - 2578

JO - Microbiology

JF - Microbiology

SN - 1350-0872

IS - 9

ER -