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Article: Blue-white selection of regulatory genes that affect the expression of dehalogenase IVa of Burkholderia cepacia MBA4

TitleBlue-white selection of regulatory genes that affect the expression of dehalogenase IVa of Burkholderia cepacia MBA4
Authors
KeywordsBurkholderia
Electro-transformation
Electroporation
Regulatory gene screening
Transposon-mutagenesis
Issue Date2007
PublisherSpringer. The Journal's web site is located at http://link.springer.de/link/service/journals/00253/index.htm
Citation
Applied Microbiology And Biotechnology, 2007, v. 76 n. 2, p. 429-437 How to Cite?
AbstractWe have developed a method for rapid screening of genes that affected the expression of dehalogenase IVa of Burkholderia cepacia MBA4. The promoter region of the dehalogenase gene was used to drive the expression of a beta-galactosidase gene. A plasmid containing this reporter was first electroporated into MBA4, and a Tn5 containing suicidal plasmid was introduced subsequently. The use of electroporation was necessary because Escherichia coli mediated transconjugation was ineffective in plasmid-carrying MBA4. The number of integrants generated was directly proportional to the amount of plasmid DNA used. Integrants with an elevated beta-galactosidase activity were isolated. Mutants with a disruption in a putative iron-transporter gene and in a putative response regulator receiver gene were identified. The basal dehalogenase transcript levels of these mutants were higher than the wild type. These mutants also grow faster than the wild type in chloroacetate-containing medium. This methodology of isolating regulatory mutants is theoretically feasible and convenient for any kinds of bacteria. © 2007 Springer-Verlag.
Persistent Identifierhttp://hdl.handle.net/10722/89232
ISSN
2015 Impact Factor: 3.376
2015 SCImago Journal Rankings: 1.262
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorFaan, YWen_HK
dc.contributor.authorYu, Men_HK
dc.contributor.authorTsang, JSHen_HK
dc.date.accessioned2010-09-06T09:54:13Z-
dc.date.available2010-09-06T09:54:13Z-
dc.date.issued2007en_HK
dc.identifier.citationApplied Microbiology And Biotechnology, 2007, v. 76 n. 2, p. 429-437en_HK
dc.identifier.issn0175-7598en_HK
dc.identifier.urihttp://hdl.handle.net/10722/89232-
dc.description.abstractWe have developed a method for rapid screening of genes that affected the expression of dehalogenase IVa of Burkholderia cepacia MBA4. The promoter region of the dehalogenase gene was used to drive the expression of a beta-galactosidase gene. A plasmid containing this reporter was first electroporated into MBA4, and a Tn5 containing suicidal plasmid was introduced subsequently. The use of electroporation was necessary because Escherichia coli mediated transconjugation was ineffective in plasmid-carrying MBA4. The number of integrants generated was directly proportional to the amount of plasmid DNA used. Integrants with an elevated beta-galactosidase activity were isolated. Mutants with a disruption in a putative iron-transporter gene and in a putative response regulator receiver gene were identified. The basal dehalogenase transcript levels of these mutants were higher than the wild type. These mutants also grow faster than the wild type in chloroacetate-containing medium. This methodology of isolating regulatory mutants is theoretically feasible and convenient for any kinds of bacteria. © 2007 Springer-Verlag.en_HK
dc.languageengen_HK
dc.publisherSpringer. The Journal's web site is located at http://link.springer.de/link/service/journals/00253/index.htmen_HK
dc.relation.ispartofApplied Microbiology and Biotechnologyen_HK
dc.subjectBurkholderiaen_HK
dc.subjectElectro-transformationen_HK
dc.subjectElectroporationen_HK
dc.subjectRegulatory gene screeningen_HK
dc.subjectTransposon-mutagenesisen_HK
dc.titleBlue-white selection of regulatory genes that affect the expression of dehalogenase IVa of Burkholderia cepacia MBA4en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0175-7598&volume=76&spage=429&epage=437&date=2007&atitle=Blue-white+selection+of+regulatory+genes+that+affect+the+expression+of+dehalogenase+IVa+of+Burkholderia+cepacia+MBA4en_HK
dc.identifier.emailTsang, JSH: jshtsang@hku.hken_HK
dc.identifier.authorityTsang, JSH=rp00792en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1007/s00253-007-1015-8en_HK
dc.identifier.pmid17530244en_HK
dc.identifier.scopuseid_2-s2.0-34547132302en_HK
dc.identifier.hkuros143116en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-34547132302&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume76en_HK
dc.identifier.issue2en_HK
dc.identifier.spage429en_HK
dc.identifier.epage437en_HK
dc.identifier.isiWOS:000248299800019-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridFaan, YW=16244861000en_HK
dc.identifier.scopusauthoridYu, M=7404272859en_HK
dc.identifier.scopusauthoridTsang, JSH=7102483508en_HK
dc.identifier.citeulike1675010-

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