Construction of improved plasmid vectors for promoter characterization in Pseudomonas aeruginosa and other gram-negative bacteria.
We report the construction of two broad host range promoter-probe plasmid vectors for rapid analysis of promoters in Gram-negative bacteria. The new vectors, pME4507 and pME4510, carry carbenicillin and gentamycin resistance genes, respectively, and are small sized (4 kb) with a flexible multiple cloning site to facilitate directional cloning of putative promoter elements. The vectors allow rapid plate-based screening for promoter activities, using beta-galactosidase as the reporter enzyme. In the absence of an inserted promoter fragment, they display very low background activity, making them a useful tool for analysis of low expression level promoters.
Rist
M
M
Institute of Microbiology, Swiss Federal Institute of Technology, ETH-Zentrum, Zürich, Switzerland.
Kertesz
M A
MA
D002228:Carbenicillin / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods; D004352:Drug Resistance, Microbial; D017930:Genes, Reporter; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D006090:Gram-Negative Bacteria / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D005839:Gentamicins; D001616:beta-Galactosidase; D002228:Carbenicillin