Activity of the hybrid trp-lac ( tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors

A broad-host-range vector, pKT240, containing the structural gene ( aph) for aminoglycoside phosphotransferase (APH), without promoter, has been constructed. Insertion of DNA fragments carrying promoters upstreamof the aph gene into the unique EcoRI site of this vector results in the expression of t...

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Veröffentlicht in:	Gene 1983-12, Vol.26 (2), p.273-282
Hauptverfasser:	Bagdasarian, Miroslawa M., Amann, Egon, Lurz, Rudolf, Rückert, Beate, Bagdasarian, Michael
Format:	Artikel
Sprache:	eng
Schlagworte:	Biological and medical sciences Biotechnology Catechol 2,3-Dioxygenase cloning vectors cloning vehicles Cloning, Molecular Dioxygenases Drug Resistance, Microbial Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Expression Regulation Genes, Bacterial Genetic engineering Genetic technics Genetic Vectors Methods. Procedures. Technologies Operon Oxygenases - genetics Plasmids promoters Pseudomonas - genetics Pseudomonas catechol 2,3-oxygenase Pseudomonas putida Recombinant DNA restriction endonucleases Streptomycin - pharmacology streptomycin resistance toluene degradation enzymes Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Zusammenfassung:	A broad-host-range vector, pKT240, containing the structural gene ( aph) for aminoglycoside phosphotransferase (APH), without promoter, has been constructed. Insertion of DNA fragments carrying promoters upstreamof the aph gene into the unique EcoRI site of this vector results in the expression of the aph gene and consequently the resistance of the host cells to streptomycin. The new vector has been used to show that the hybrid trp-lac ( tac) promoter and the promoter of the lacI Q gene of Escherichia coli are active in Pseudomonas putida. Derivatives of pKT240 containing tac and lacI Q sequences may be used as wide-host-range expression vectors. Regulated overproduction of APH and catechol 2,3-oxygenase can be obtained with the aid of the new vectors in both E. coli and P. putida.
ISSN:	0378-1119 1879-0038
DOI:	10.1016/0378-1119(83)90197-X