A small derivative of the broad-host-range plasmid RK2 which can
be switched from a replicating to a non-replicating state as a response
to an externally added inducer.
TrfA is the only plasmid-encoded protein required for RK2 replication.
We report here the construction and characterization of an RK2-based
vector in which trfA is expressed from the inducible promoter Pm.
The resulting construct, pJBSD1, was found to replicate in Escherichia
coli DH5a (recA(-)) only in the presence of a Pm inducer. In two
tested E. coli recA(+) strains pJBSD1 could replicate in the absence
of inducer, but a replication inducer-dependent phenotype was obtained
in these strains by introducing a mutation known to reduce the trfA
expression level. The plasmid construct could be used as a conditional
suicide vector system for targeted chromosomal integration via homologous
recombination. This feature may potentially be used for many types
of studies in microbial molecular biology.
%0 Journal Article
%1 Karunakaran1999
%A Karunakaran, P.
%A Endresen, D. T.
%A Ertesvag, H.
%A Blatny, J. M.
%A Valla, S.
%D 1999
%J FEMS Microbiol. Lett.
%K & (Genetics) *DNA *Escherichia *Gene *Genetic ; Bacterial Chain Escherichia Expression Gov't Non-U.S. Plasmids/*genetics Polymerase Promoter Proteins Proteins/genetics/metabolism Reaction Regions Regulation, Replication Research Support, Temperature Transformation, Vectors coli coli/*genetics/growth development
%N 2
%P 221-7
%T A small derivative of the broad-host-range plasmid RK2 which can
be switched from a replicating to a non-replicating state as a response
to an externally added inducer.
%U http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?cmd=prlinks&dbfrom=pubmed&retmode=ref&id=10556715
%V 180
%X TrfA is the only plasmid-encoded protein required for RK2 replication.
We report here the construction and characterization of an RK2-based
vector in which trfA is expressed from the inducible promoter Pm.
The resulting construct, pJBSD1, was found to replicate in Escherichia
coli DH5a (recA(-)) only in the presence of a Pm inducer. In two
tested E. coli recA(+) strains pJBSD1 could replicate in the absence
of inducer, but a replication inducer-dependent phenotype was obtained
in these strains by introducing a mutation known to reduce the trfA
expression level. The plasmid construct could be used as a conditional
suicide vector system for targeted chromosomal integration via homologous
recombination. This feature may potentially be used for many types
of studies in microbial molecular biology.
@article{Karunakaran1999,
__markedentry = {[phpts:6]},
abstract = {TrfA is the only plasmid-encoded protein required for RK2 replication.
We report here the construction and characterization of an RK2-based
vector in which trfA is expressed from the inducible promoter Pm.
The resulting construct, pJBSD1, was found to replicate in Escherichia
coli DH5a (recA(-)) only in the presence of a Pm inducer. In two
tested E. coli recA(+) strains pJBSD1 could replicate in the absence
of inducer, but a replication inducer-dependent phenotype was obtained
in these strains by introducing a mutation known to reduce the trfA
expression level. The plasmid construct could be used as a conditional
suicide vector system for targeted chromosomal integration via homologous
recombination. This feature may potentially be used for many types
of studies in microbial molecular biology.},
added-at = {2011-11-04T13:47:04.000+0100},
author = {Karunakaran, P. and Endresen, D. T. and Ertesvag, H. and Blatny, J. M. and Valla, S.},
authoraddress = {UNIGEN Center for Molecular Biology and Department of Biotechnology,
Norwegian University of Science and Technology, 7489, Trondheim,
Norway.},
biburl = {https://www.bibsonomy.org/bibtex/273ce3c6a7c33283ef0681b020ae8d607/pawelsikorski},
interhash = {a849a15b14a91b6398ad71bb423158c3},
intrahash = {73ce3c6a7c33283ef0681b020ae8d607},
journal = {FEMS Microbiol. Lett.},
keywords = {& (Genetics) *DNA *Escherichia *Gene *Genetic ; Bacterial Chain Escherichia Expression Gov't Non-U.S. Plasmids/*genetics Polymerase Promoter Proteins Proteins/genetics/metabolism Reaction Regions Regulation, Replication Research Support, Temperature Transformation, Vectors coli coli/*genetics/growth development},
language = {eng},
medline-aid = {S0378-1097(99)00476-0 [pii]},
medline-da = {20000111},
medline-dcom = {20000111},
medline-edat = {1999/11/11},
medline-fau = {Karunakaran, P ; Endresen, D T ; Ertesvag, H ; Blatny, J M ; Valla,
S},
medline-is = {0378-1097 (Print)},
medline-jid = {7705721},
medline-jt = {FEMS microbiology letters.},
medline-lr = {20060501},
medline-mhda = {1999/11/11 00:01},
medline-own = {NLM},
medline-pl = {NETHERLANDS},
medline-pmid = {10556715},
medline-pst = {ppublish},
medline-pt = {Journal Article},
medline-pubm = {Print},
medline-rn = {0 (Bacterial Proteins) ; 0 (Escherichia coli Proteins) ; 0 (TrfA protein,
E coli)},
medline-sb = {IM},
medline-so = {FEMS Microbiol Lett. 1999 Nov 15;180(2):221-7.},
medline-stat = {MEDLINE},
number = 2,
owner = {phpts},
pages = {221-7},
timestamp = {2011-11-04T13:47:14.000+0100},
title = {A small derivative of the broad-host-range plasmid RK2 which can
be switched from a replicating to a non-replicating state as a response
to an externally added inducer.},
url = {http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?cmd=prlinks\&dbfrom=pubmed\&retmode=ref\&id=10556715},
volume = 180,
year = 1999
}