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Development of a gene expression vector for Thermus thermophilus based on the promoter of the respiratory nitrate reductase
Centro de Biología Molecular 'Severo Ochoa' CSIC-UAM, Campus de Cantoblanco, Madrid, Spain.
Centro de Biología Molecular 'Severo Ochoa' CSIC-UAM, Campus de Cantoblanco, Madrid, Spain.
Centro de Biología Molecular 'Severo Ochoa' CSIC-UAM, Campus de Cantoblanco, Madrid, Spain.
Centro de Biología Molecular 'Severo Ochoa' CSIC-UAM, Campus de Cantoblanco, Madrid, Spain.
2003 (English)In: Plasmid, ISSN 0147-619X, E-ISSN 1095-9890, Vol. 49, no 1, 2-8 p.Article in journal (Refereed) Published
Abstract [en]

A specific expression system for Thermus spp. is described. Plasmid pMKE1 contains replicative origins for Escherichia coli and Thermus spp., a selection gene encoding a thermostable resistance to kanamycin, and a 720 bp DNA region containing the promoter (Pnar), and the regulatory sequences of the respiratory nitrate reductase operon of Thermus thermophilus HB8. Two genes, encoding a thermophilic beta-galactosidase and an alkaline phosphatase were cloned in pMKE1 as cytoplasmic and periplasmic reporters, respectively. The expression of the reporters was specifically induced by the combined action of nitrate and anoxia in facultative anaerobic derivatives of T. thermophilus HB27 to which the gene cluster for nitrate respiration was transferred by conjugation. Overexpressions in the range of approximately 200-fold were obtained for the cytoplasmic reporter, whereas that of the periplasmic reporter was limited to approximately 20-fold, with respect to their intrinsic respective activities.

Place, publisher, year, edition, pages
2003. Vol. 49, no 1, 2-8 p.
Keyword [en]
thermophilic, vector, expression, Thermus, reporter
National Category
Genetics
Identifiers
URN: urn:nbn:se:umu:diva-81878DOI: 10.1016/S0147-619X(02)00146-4ISI: 000181264300001PubMedID: 12583995OAI: oai:DiVA.org:umu-81878DiVA: diva2:658626
Available from: 2013-10-22 Created: 2013-10-22 Last updated: 2017-12-06Bibliographically approved

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CiteExportLink to record
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