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PMID: 3039307 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The origin of transfer (oriT) of the conjugative plasmid R46: characterization by deletion analysis and DNA sequencing.

Molecular & general genetics : MGG ·Vol. 208 ·No. 1-2 ·1987-06-00 ·Pages 219-25

Coupland GM, Brown AM, Willetts NS

Abstract

The origin of transfer (oriT) is the sequence within which conjugal transfer of plasmid DNA is initiated, and is absolutely required in cis for plasmid mobilization. We have cloned oriT from the 52 kb IncN plasmid R46 on a 600 bp fragment, and mapped the limits of the relevant sequence by deletion analysis and transposon mutagenesis. The nucleotide sequence of the oriT region contains 13 direct repeats of an 11 bp consensus sequence, 3 different pairs of 10 bp inverted repeats, and a segment that is extremely A-T rich. The direct repeats are within a region required for high frequency transfer and their sequence is such that their periodic alignment along the helix may induce curvature of the DNA. Analysis of Tn1725 insertions within the sequenced fragment of R46 revealed that, unlike most other transposons, transposition of Tn1725 can cause target sequence duplications of three different sizes.

MeSH Terms
Base Sequence Chromosome Deletion Cloning, Molecular Conjugation, Genetic DNA Replication DNA Transposable Elements Escherichia coli/genetics Genes, Bacterial Mutation Plasmids Promoter Regions, Genetic
Chemicals
DNA Transposable Elements
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Coupland G M
Brown A M
Willetts N S
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41 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00330445
Published
1987-06-00
Pages
219-25
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00330445
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