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dc.creatorNikodinović-Runić, Jasmina
dc.creatorPriestley, Nigel D.
dc.date.accessioned2018-11-22T00:09:32Z
dc.date.available2018-11-22T00:09:32Z
dc.date.issued2006
dc.identifier.issn0147-619X
dc.identifier.urihttp://cherry.chem.bg.ac.rs/handle/123456789/764
dc.description.abstractAn Escherichia coli-Streptomyces shuttle vector (pJN100) was constructed, by inserting an origin of transfer (oriT), derived from the E coli broad host range plasmid RK2, into pANT1202, a high-copy-number vector for gene expression in Streptomyces. The resulting conjugably transferable vector contains the pANT1202-derived SnpR (LysR-like protein) activated snpA promoter that drives strong heterologous expression of proteins. We initially demonstrated that plasmid pJN 100 was transferred with high frequency (10(-5-7) exconjugants per recipient) into several Streptomyces strains that were refractory to transformation by other means. Plasmid pJN100 was also shown to be stable in E. coli and Streptomyces. We confirmed functional protein expression by using a pJN100 derivative to complement a mutant of Streptomyces griseus with a disrupted chromosomal copy of the gene nonM, a gene encoding an essential reductase in the nonactin biosynthesis gene cluster. High levels of protein expression were confirmed using Western blotting to assess the production of the serine esterase NonR, an enzyme responsible for nonactin resistance in the nonactin producer S. griseus. (c) 2006 Elsevier Inc. All rights reserved.en
dc.publisherAcademic Press Inc Elsevier Science, San Diego
dc.rightsrestrictedAccess
dc.sourcePlasmid
dc.subjectStreptomycesen
dc.subjectconjugationen
dc.subjectshuttle vectoren
dc.subjectoriTen
dc.subjectsnpA promoteren
dc.subjectRK2en
dc.subjectheterologous expressionen
dc.titleA second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugationen
dc.typearticle
dc.rights.licenseARR
dcterms.abstractНикодиновић-Рунић, Јасмина; Приестлеy, Нигел Д.;
dc.citation.volume56
dc.citation.issue3
dc.citation.spage223
dc.citation.epage227
dc.identifier.wos000242306000008
dc.identifier.doi10.1016/j.plasmid.2006.05.002
dc.citation.other56(3): 223-227
dc.citation.rankM23
dc.identifier.pmid16806469
dc.type.versionpublishedVersionen
dc.identifier.scopus2-s2.0-33750453762
dc.identifier.rcubKon_1717


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