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

doi:10.1016/S0378-1097(03)00171-X

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 journal › Article › peer-review

19 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 language	English
Pages (from-to)	125-130
Number of pages	6
Journal	FEMS microbiology letters
Volume	221
Issue number	1
DOIs	https://doi.org/10.1016/S0378-1097(03)00171-X
Publication status	Published - 2003 Apr 11
Externally published	Yes

Keywords

GFPuv
Gene expression
Promoter
Ribosome-binding site
TA-cloning

ASJC Scopus subject areas

Microbiology
Molecular Biology
Genetics

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10.1016/S0378-1097(03)00171-X

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

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.",

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author = "Yoshiaki Ohashi and Hideyuki Ohshima and Kenji Tsuge and Mitsuhiro Itaya",

note = "Funding Information: We are grateful to Q. Hua and K. Shimizu for technical advice on the fluorescence measurements. This work was supported in part by a grant from the New Energy and Industrial Technology Development Organization (NEDO) of the Ministry of Economy, Trade and Industry of Japan (Development of a Technological Infrastructure for Industrial Bioprocesses Project).",

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AU - Ohashi, Yoshiaki

AU - Ohshima, Hideyuki

AU - Tsuge, Kenji

AU - Itaya, Mitsuhiro

N1 - Funding Information: We are grateful to Q. Hua and K. Shimizu for technical advice on the fluorescence measurements. This work was supported in part by a grant from the New Energy and Industrial Technology Development Organization (NEDO) of the Ministry of Economy, Trade and Industry of Japan (Development of a Technological Infrastructure for Industrial Bioprocesses Project).

PY - 2003/4/11

Y1 - 2003/4/11

N2 - 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.

AB - 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.

KW - GFPuv

KW - Gene expression

KW - Promoter

KW - Ribosome-binding site

KW - TA-cloning

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

Abstract

Keywords

ASJC Scopus subject areas

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