Construction of broad-host-range vectors for the selection of divergent promoters

• Published in: Gene Vol. 90; no. 1; pp. 145 - 148
• Main Authors: Ronald, S.L., Kropinski, A.M., Farinha, M.A.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 31.05.1990; Amsterdam Elsevier; New York, NY
• Subjects: alkaline phosphatase; Biological and medical sciences; Biotechnology; Cloning, Molecular; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Genetic Vectors; luciferase; Methods. Procedures. Technologies; multiple cloning site; phage D3; Plasmids; plasmids pBR322 and pRO1614; promoter; Promoter Regions, Genetic; Pseudomonas - genetics; Pseudomonas aeruginosa; Recombinant DNA; reporter gene; Restriction Mapping; tetradecanal; β-galactosidase; phage D3; tsc; Escherichia coli; nt; tetA; Recombinant DNA; bp; Tc; plasmids pBR322 and pRO1614; β-galactosidase; SD; bla; cat; kb; luxAB; Pseudomonas aeruginosa; promoter; alkaline phosphatase; reporter gene; phoA; TSB; ori; multiple cloning site; Lux; AP; βGal; p; XGal; luciferase; XP; tetR; MCS; tetradecanal; lacZ; Cb; Reporter gene; Transcription promoter; DNA; Genetic engineering; Molecular cloning; Gene expression; Vector
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(90)90451-V

A series of promoter-probe plasmid vectors has been constructed which allows for the selection of DNA sequences containing divergent control elements. Each vector contains a pair of promoterless genes [encoding β-galactosidase ( lacZ), alkaline phosphatase ( phoA), and bacterial luciferase ( luxAB)] arranged in an antiparallel fashion and separated by a large intervening multiple cloning site. The vectors permit direct detection of promoter activity on indicator plates after transformation. Cloned promoters are selected based on production of coloured products in the case of lacZ and phoA, and by the emission of light in the case of luxAB. These vectors have been tested using known divergent promoter elements from pBR322 and Pseudomonas phage D3.