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Article: Use of ribosomal promoters from Burkholderia cenocepacia and Burkholderia cepacia for improved expression of transporter protein in Escherichia coli

TitleUse of ribosomal promoters from Burkholderia cenocepacia and Burkholderia cepacia for improved expression of transporter protein in Escherichia coli
Authors
KeywordsBurkholderia spp.
Haloacetate permease
Monochloroacetate
Ribosomal promoter
Transporter protein
Issue Date2006
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprep
Citation
Protein Expression And Purification, 2006, v. 49 n. 2, p. 219-227 How to Cite?
AbstractExpression of heterologous protein in Escherichia coli usually based on the IPTG-inducible expression systems. The use of these systems for membrane protein production, however, usually caused cytotoxic problem that affected the yield and functional characterization of the protein. Optimization of these systems for transporter protein production is time-consuming and is usually ineffective. Here, we described the use of the ribosomal promoters Ps12 from Burkholderia cenocepacia LMG16656 and from Burkholderia cepacia MBA4 for efficient expression of functional transporter protein in E. coli. These promoters were used to drive the expression of a transmembrane protein, Deh4p, which help transport monohaloacetates into B. cepacia MBA4 for metabolism. Production of Deh4p in E. coli using an IPTG-inducible promoter resulted in no expression in uninduced condition and cell lysis in the presence of IPTG. Moreover, it has been reported that IPTG increased the endogenous production of other permeases such as LacZ and MelB. Cells expressing Deh4p from a Ps12 promoter grew normally in rich medium and which did not increase the expression of other permease. Uptake of 14C-monochloroacetic acid has confirmed the production of the transporter protein in these cells. The results showed that the constitutive ribosomal protein promoters from the Burkholderia sp. could be used for effective expression of transporter protein in E. coli without causing any detrimental and unnecessary effect. © 2006 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/68466
ISSN
2015 Impact Factor: 1.407
2015 SCImago Journal Rankings: 0.767
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYu, Men_HK
dc.contributor.authorTsang, JSHen_HK
dc.date.accessioned2010-09-06T06:04:51Z-
dc.date.available2010-09-06T06:04:51Z-
dc.date.issued2006en_HK
dc.identifier.citationProtein Expression And Purification, 2006, v. 49 n. 2, p. 219-227en_HK
dc.identifier.issn1046-5928en_HK
dc.identifier.urihttp://hdl.handle.net/10722/68466-
dc.description.abstractExpression of heterologous protein in Escherichia coli usually based on the IPTG-inducible expression systems. The use of these systems for membrane protein production, however, usually caused cytotoxic problem that affected the yield and functional characterization of the protein. Optimization of these systems for transporter protein production is time-consuming and is usually ineffective. Here, we described the use of the ribosomal promoters Ps12 from Burkholderia cenocepacia LMG16656 and from Burkholderia cepacia MBA4 for efficient expression of functional transporter protein in E. coli. These promoters were used to drive the expression of a transmembrane protein, Deh4p, which help transport monohaloacetates into B. cepacia MBA4 for metabolism. Production of Deh4p in E. coli using an IPTG-inducible promoter resulted in no expression in uninduced condition and cell lysis in the presence of IPTG. Moreover, it has been reported that IPTG increased the endogenous production of other permeases such as LacZ and MelB. Cells expressing Deh4p from a Ps12 promoter grew normally in rich medium and which did not increase the expression of other permease. Uptake of 14C-monochloroacetic acid has confirmed the production of the transporter protein in these cells. The results showed that the constitutive ribosomal protein promoters from the Burkholderia sp. could be used for effective expression of transporter protein in E. coli without causing any detrimental and unnecessary effect. © 2006 Elsevier Inc. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprepen_HK
dc.relation.ispartofProtein Expression and Purificationen_HK
dc.subjectBurkholderia spp.en_HK
dc.subjectHaloacetate permeaseen_HK
dc.subjectMonochloroacetateen_HK
dc.subjectRibosomal promoteren_HK
dc.subjectTransporter proteinen_HK
dc.titleUse of ribosomal promoters from Burkholderia cenocepacia and Burkholderia cepacia for improved expression of transporter protein in Escherichia colien_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1046-5928&volume=49&spage=219&epage=227&date=2006&atitle=Use+of+ribosomal+promoters+from+Burkholderia+cenocepacia+and+Burkholderia+cepacia+for+improved+expression+of+transporter+protein+in+Escherichia+colien_HK
dc.identifier.emailTsang, JSH: jshtsang@hku.hken_HK
dc.identifier.authorityTsang, JSH=rp00792en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.pep.2006.04.004en_HK
dc.identifier.pmid16737826-
dc.identifier.scopuseid_2-s2.0-33748785968en_HK
dc.identifier.hkuros124640en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33748785968&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume49en_HK
dc.identifier.issue2en_HK
dc.identifier.spage219en_HK
dc.identifier.epage227en_HK
dc.identifier.isiWOS:000241206000010-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridYu, M=7404272859en_HK
dc.identifier.scopusauthoridTsang, JSH=7102483508en_HK

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