Characterization of the ColE1 mobilization region and its protein products

A. Christopher Boyd; John A.K. Archer; David J. Sherratt

doi:10.1007/BF02464922

Characterization of the ColE1 mobilization region and its protein products

A. Christopher Boyd^*, John A.K. Archer, David J. Sherratt

^*Corresponding author for this work

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

63 Scopus citations

Abstract

A third of the 6.6 kb genome of ColE1 is devoted to mobilization (mob) genes necessary to promote its specific transfer in the presence of conjugative plasmids. The mob region is genetically complex: two mob genes are entirely overlapped by a third. Oligonucleotide-directed mutagenesis was used to insert an amber codon into one of the overlapped genes and make possible a full complementation analysis of mob. Four mob genes essential for mobilization by R64 drd11 were thus identified. Fragments of mob were subcloned under control of the P_tac promoter in a suitable vector, overexpressed in minicells and the mobilization proteins visualized. A comprehensive alignment of the mob region of ColE1 with those of its close relatives ColK and ColA demonstrating that the four essential mob genes are conserved is also presented.

Original language	English (US)
Pages (from-to)	488-498
Number of pages	11
Journal	MGG Molecular & General Genetics
Volume	217
Issue number	2-3
DOIs	https://doi.org/10.1007/BF02464922
State	Published - Jun 1989
Externally published	Yes

Keywords

ColE1
Overlapping genes
Plasmid mobilization
Relaxation complex
Site-directed mutagenesis

ASJC Scopus subject areas

Genetics

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10.1007/BF02464922

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title = "Characterization of the ColE1 mobilization region and its protein products",

abstract = "A third of the 6.6 kb genome of ColE1 is devoted to mobilization (mob) genes necessary to promote its specific transfer in the presence of conjugative plasmids. The mob region is genetically complex: two mob genes are entirely overlapped by a third. Oligonucleotide-directed mutagenesis was used to insert an amber codon into one of the overlapped genes and make possible a full complementation analysis of mob. Four mob genes essential for mobilization by R64 drd11 were thus identified. Fragments of mob were subcloned under control of the Ptac promoter in a suitable vector, overexpressed in minicells and the mobilization proteins visualized. A comprehensive alignment of the mob region of ColE1 with those of its close relatives ColK and ColA demonstrating that the four essential mob genes are conserved is also presented.",

keywords = "ColE1, Overlapping genes, Plasmid mobilization, Relaxation complex, Site-directed mutagenesis",

author = "{Christopher Boyd}, A. and Archer, {John A.K.} and Sherratt, {David J.}",

year = "1989",

month = jun,

doi = "10.1007/BF02464922",

language = "English (US)",

volume = "217",

pages = "488--498",

journal = "MGG Molecular & General Genetics",

issn = "0026-8925",

publisher = "Springer Verlag",

number = "2-3",

}

TY - JOUR

T1 - Characterization of the ColE1 mobilization region and its protein products

AU - Christopher Boyd, A.

AU - Archer, John A.K.

AU - Sherratt, David J.

PY - 1989/6

Y1 - 1989/6

N2 - A third of the 6.6 kb genome of ColE1 is devoted to mobilization (mob) genes necessary to promote its specific transfer in the presence of conjugative plasmids. The mob region is genetically complex: two mob genes are entirely overlapped by a third. Oligonucleotide-directed mutagenesis was used to insert an amber codon into one of the overlapped genes and make possible a full complementation analysis of mob. Four mob genes essential for mobilization by R64 drd11 were thus identified. Fragments of mob were subcloned under control of the Ptac promoter in a suitable vector, overexpressed in minicells and the mobilization proteins visualized. A comprehensive alignment of the mob region of ColE1 with those of its close relatives ColK and ColA demonstrating that the four essential mob genes are conserved is also presented.

AB - A third of the 6.6 kb genome of ColE1 is devoted to mobilization (mob) genes necessary to promote its specific transfer in the presence of conjugative plasmids. The mob region is genetically complex: two mob genes are entirely overlapped by a third. Oligonucleotide-directed mutagenesis was used to insert an amber codon into one of the overlapped genes and make possible a full complementation analysis of mob. Four mob genes essential for mobilization by R64 drd11 were thus identified. Fragments of mob were subcloned under control of the Ptac promoter in a suitable vector, overexpressed in minicells and the mobilization proteins visualized. A comprehensive alignment of the mob region of ColE1 with those of its close relatives ColK and ColA demonstrating that the four essential mob genes are conserved is also presented.

KW - ColE1

KW - Overlapping genes

KW - Plasmid mobilization

KW - Relaxation complex

KW - Site-directed mutagenesis

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U2 - 10.1007/BF02464922

DO - 10.1007/BF02464922

M3 - Article

C2 - 2671664

AN - SCOPUS:0024677442

SN - 0026-8925

VL - 217

SP - 488

EP - 498

JO - MGG Molecular & General Genetics

JF - MGG Molecular & General Genetics

IS - 2-3

ER -

Characterization of the ColE1 mobilization region and its protein products

Abstract

Keywords

ASJC Scopus subject areas

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