%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 }