Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa

Herbert P. Schweizer

doi:10.1016/0378-1119(91)90396-S

Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa

Herbert P. Schweizer

Research output: Contribution to journal › Article › peer-review

44 Scopus citations

Abstract

Several new broad-host-range vectors for the construction of protein fusions to the Escherichia coli lacZ gene have been developed. In all of the constructs, a multiple cloning site (MCS) containing unique restriction sites is located upstream of lac operon segments whose lacZ genes lack translational start signals. Some of the vectors (pPZ10, pPZ20 and pPZ30) also contain transcriptional terminators upstream of the MCS. The new vectors allow the fusion of genes to lacZ in all translational reading frames. Due to a higher copy number they allow direct screening in E. coli for weakly expressed foreign promoters. Their usefulness for gene analysis in Pseudomonas aemginosa was demonstrated by construction and expression of a regA'::'lac Z-encoded protein fusion.

Original language	English (US)
Pages (from-to)	87-92
Number of pages	6
Journal	Gene
Volume	103
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(91)90396-S
State	Published - Jul 15 1991
Externally published	Yes

Keywords

Gram-negative
M13 bacteriophage
Recombinant DNA
gene fusion
promoter detection
reading frame
transcription
translation
β-galactosidase

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(91)90396-S

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title = "Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa",

abstract = "Several new broad-host-range vectors for the construction of protein fusions to the Escherichia coli lacZ gene have been developed. In all of the constructs, a multiple cloning site (MCS) containing unique restriction sites is located upstream of lac operon segments whose lacZ genes lack translational start signals. Some of the vectors (pPZ10, pPZ20 and pPZ30) also contain transcriptional terminators upstream of the MCS. The new vectors allow the fusion of genes to lacZ in all translational reading frames. Due to a higher copy number they allow direct screening in E. coli for weakly expressed foreign promoters. Their usefulness for gene analysis in Pseudomonas aemginosa was demonstrated by construction and expression of a regA'::'lac Z-encoded protein fusion.",

keywords = "Gram-negative, M13 bacteriophage, Recombinant DNA, gene fusion, promoter detection, reading frame, transcription, translation, β-galactosidase",

author = "Schweizer, {Herbert P.}",

note = "Funding Information: This researchw as supported by grants from the Medical Research Council of Canada, an institutional grant from the University of Calgary, and a Ruth Rannie Memorial Endowment grant from the Faculty of Medicine at the University of Calgary. H.P.S. is a MRC Medical Scholar. I wish to thank D. Storey for the gift of clone pDF202 and critical reading of the manuscript, and W. Boos for providing the pNM plasmids. I also wish to thank A. Schryvers for sharing laboratory facilities.",

year = "1991",

month = jul,

day = "15",

doi = "10.1016/0378-1119(91)90396-S",

language = "English (US)",

volume = "103",

pages = "87--92",

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T1 - Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa

AU - Schweizer, Herbert P.

N1 - Funding Information: This researchw as supported by grants from the Medical Research Council of Canada, an institutional grant from the University of Calgary, and a Ruth Rannie Memorial Endowment grant from the Faculty of Medicine at the University of Calgary. H.P.S. is a MRC Medical Scholar. I wish to thank D. Storey for the gift of clone pDF202 and critical reading of the manuscript, and W. Boos for providing the pNM plasmids. I also wish to thank A. Schryvers for sharing laboratory facilities.

PY - 1991/7/15

Y1 - 1991/7/15

N2 - Several new broad-host-range vectors for the construction of protein fusions to the Escherichia coli lacZ gene have been developed. In all of the constructs, a multiple cloning site (MCS) containing unique restriction sites is located upstream of lac operon segments whose lacZ genes lack translational start signals. Some of the vectors (pPZ10, pPZ20 and pPZ30) also contain transcriptional terminators upstream of the MCS. The new vectors allow the fusion of genes to lacZ in all translational reading frames. Due to a higher copy number they allow direct screening in E. coli for weakly expressed foreign promoters. Their usefulness for gene analysis in Pseudomonas aemginosa was demonstrated by construction and expression of a regA'::'lac Z-encoded protein fusion.

AB - Several new broad-host-range vectors for the construction of protein fusions to the Escherichia coli lacZ gene have been developed. In all of the constructs, a multiple cloning site (MCS) containing unique restriction sites is located upstream of lac operon segments whose lacZ genes lack translational start signals. Some of the vectors (pPZ10, pPZ20 and pPZ30) also contain transcriptional terminators upstream of the MCS. The new vectors allow the fusion of genes to lacZ in all translational reading frames. Due to a higher copy number they allow direct screening in E. coli for weakly expressed foreign promoters. Their usefulness for gene analysis in Pseudomonas aemginosa was demonstrated by construction and expression of a regA'::'lac Z-encoded protein fusion.

KW - Gram-negative

KW - M13 bacteriophage

KW - Recombinant DNA

KW - gene fusion

KW - promoter detection

KW - reading frame

KW - transcription

KW - translation

KW - β-galactosidase

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U2 - 10.1016/0378-1119(91)90396-S

DO - 10.1016/0378-1119(91)90396-S

M3 - Article

C2 - 1908810

SN - 0378-1119

VL - 103

SP - 87

EP - 92

JO - Gene

JF - Gene

IS - 1

ER -

Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa

Abstract

Keywords

ASJC Scopus subject areas

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