A set of cassettes and improved vectors for genetic and biochemical characterization of pseudomonas genes

• Published in: Gene Vol. 57; no. 1; pp. 61 - 72
• Main Authors: Deretic, V., Chandrasekharappa, S., Gill, J.F., Chatterjee, D.K., Chakrabarty, A.M.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 1987; Amsterdam Elsevier; New York, NY
• Subjects: Applied microbiology; Biological and medical sciences; Biotechnology; Cloning, Molecular; Coliphages - genetics; DNA, Recombinant - metabolism; Escherichia coli - genetics; expression vectors; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic engineering; Genetic technics; Genetic Vectors; maxicell analysis; Methods. Procedures. Technologies; Microbiology; Miscellaneous; Plasmids; Protein Biosynthesis; Pseudomonas; Pseudomonas - genetics; Pseudomonas aeruginosa - genetics; Pseudomonas gene cloning; Recombinant DNA; Recombinant Fusion Proteins - analysis; Vectors (cloning, transfer, expression). Insertion sequences and transposons; xylE translational fusions; Km; ORF; Pseudomonas gene cloning; mU; Recombinant DNA; bp; Ap; βGal; CDO; Tc; xylE translational fusions; r; XGal; maxicell analysis; Δ; PolIk; kb; IPTG; Sm; expression vectors; alg; GMD; lacZ′ or lacZα; Pseudomonadales; Gene; Bacteria; Pseudomonadaceae; Biosynthesis; Alginates; Molecular cloning; Gene expression; Pseudomonas; Vector
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(87)90177-6

A set of broad-host-range vectors allowing direct selection of recombinant DNA molecules to facilitate subcloning and expression analyses of Pseudomonas genes was constructed using BglII lacZα cassette. Controlled expression vectors pVDtac39 and pVDtac24 were shown to be useful for determination of enzymatic activities encoded by the cloned DNA fragments and M r determination of the corresponding polypeptides. A set of Pseudomonas putida xylE gene cassettes truncated at the 5' end was constructed for translational (protein) fusion studies. A protein fusion of the Pseudomonas aeruginosa algD gene, coding for GDPmannose dehydro-genase, and the truncated xylE gene cassette was used to verify the putative coding region and translational signals predicted from the algD nucleotide sequence.