Far different levels of gene expression provided by an oriented cloning system in Bacillus subtilis and Escherichia coli

Yoshiaki Ohashi, Hideyuki Ohshima, Kenji Tsuge, Mitsuhiro Itaya

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

A gene expression system for both Bacillus subtilis and Escherichia coli was developed. The expression vector, pHASH102, produces any combination of promoter and open reading frame to be expressed based on the T-extended cloning method. Because the pHASH series vectors are designed to shuttle between the genome and a high copy plasmid in B. subtilis, the expression profiles of copy number dependence can be examined systematically. We demonstrated that vectors with Pr, Pspac, and PS10 promoters are suitable for the overexpression of GFPuv. Moreover, aadK encoding aminoglycoside 6-adenylyltransferase (a streptomycin-resistance gene) of B. subtilis was successfully overexpressed in both B. subtilis and E. coli. These highly expressed GFPuv and aadK genes can be used as a genetic marker for both organisms.

Original languageEnglish
Pages (from-to)125-130
Number of pages6
JournalFEMS Microbiology Letters
Volume221
Issue number1
DOIs
Publication statusPublished - 2003 Apr 11

Fingerprint

Bacillus subtilis
Organism Cloning
Escherichia coli
Gene Expression
Streptomycin
Genetic Markers
Open Reading Frames
Genes
Plasmids
Genome

Keywords

  • Gene expression
  • GFPuv
  • Promoter
  • Ribosome-binding site
  • TA-cloning

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Far different levels of gene expression provided by an oriented cloning system in Bacillus subtilis and Escherichia coli. / Ohashi, Yoshiaki; Ohshima, Hideyuki; Tsuge, Kenji; Itaya, Mitsuhiro.

In: FEMS Microbiology Letters, Vol. 221, No. 1, 11.04.2003, p. 125-130.

Research output: Contribution to journalArticle

Ohashi, Yoshiaki ; Ohshima, Hideyuki ; Tsuge, Kenji ; Itaya, Mitsuhiro. / Far different levels of gene expression provided by an oriented cloning system in Bacillus subtilis and Escherichia coli. In: FEMS Microbiology Letters. 2003 ; Vol. 221, No. 1. pp. 125-130.
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