Comparison of the CopB systems of plasmids R1 and Co1V2-K94: A single base alteration in CopB gene is responsible for the increased copy number of the low copy number plasmid CoIV2-K94

SUMMARYThe insertion of a high-copy-number plasmid into a lambdoid phage chromosome which lacks a functional repressor gene confers on the hybrid ‘phasmid’ the capacity to grow on an immune lysogen. This was found to be due to titration of repressor because of plasmid replication. We have exploited this property in order to isolate mutants that affect plasmid replication. These mutants have been mapped in a region that was previously characterized as necessary for plasmid replication and incompatibility properties. Some of the mutations could revert at frequencies characteristic of single-base-pair change mutations.

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Detection of displacement (“D”) loops with the properties of a replicating intermediate synthesized by a DNA/membrane complex derived from the low-copy-number plasmid RK2

Plasmid ◽

10.1016/0147-619x(84)90051-9 ◽

1984 ◽

Vol 12 (3) ◽

pp. 227-232 ◽

Cited By ~ 2

Author(s):

William Firshein ◽

Lucien Caro

Keyword(s):

Copy Number ◽

Membrane Complex ◽

Copy Number Plasmid ◽

Low Copy Number ◽

Plasmid Rk2

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Targeted Isolation of a Designated Region of the Bacillus subtilis Genome by Recombinational Transfer

Applied and Environmental Microbiology ◽

10.1128/aem.70.4.2508-2513.2004 ◽

2004 ◽

Vol 70 (4) ◽

pp. 2508-2513 ◽

Cited By ~ 12

Author(s):

Satoshi Tomita ◽

Kenji Tsuge ◽

Yo Kikuchi ◽

Mitsuhiro Itaya

Keyword(s):

Bacillus Subtilis ◽

Copy Number ◽

Positional Cloning ◽

Deletion Mutant ◽

Essential Gene ◽

Transfer System ◽

Copy Number Plasmid ◽

Low Copy Number ◽

Subtilis Genome

ABSTRACT A method for positional cloning of the Bacillus subtilis genome was developed. The method requires a set of two small DNA fragments that flank the region to be copied. A 38-kb segment that carries genes ppsABCDE encoding five enzymes for antibiotic plipastatin synthesis and another genome locus as large as 100 kb including one essential gene were examined for positional cloning. The positional cloning vector for ppsABCDE was constructed using a B. subtilis low-copy-number plasmid that faithfully copied the precise length of the 38-kb DNA in vivo via the recombinational transfer system of this bacterium. Structure of the copied DNA was confirmed by restriction enzyme analyses. Furthermore, the unaltered structure of the 38-kb DNA was demonstrated by complementation of a ppsABCDE deletion mutant.

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A vector for studying plasmid stability functions in Streptomyces

Genetics Research ◽

10.1017/s0016672300023971 ◽

1988 ◽

Vol 51 (1) ◽

pp. 71-74 ◽

Cited By ~ 4

Author(s):

Kevin Kendall ◽

John Cullum

Keyword(s):

Copy Number ◽

Stability Region ◽

Selectable Marker ◽

Plasmid Stability ◽

Stability Function ◽

Melanin Production ◽

Stability Functions ◽

Tyrosinase Gene ◽

Copy Number Plasmid ◽

Low Copy Number

SummaryWe constructed a cloning vector (pMT603) based on the low copy number plasmid SCP2*. pMT6O3 is unstable because it lacks the SCP2* stability region and carries the selectable marker thiostrepton-resistance and a tyrosinase gene which results in melanin production. This allows easy testing of plasmid stability and we demonstrated its usefulness by cloning a plasmid stability function.

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