Characterization of the phospholipase C gene of Pseudomonas aeruginosa cloned in Escherichia coli |
| |
Authors: | S Lory P C Tai |
| |
Affiliation: | Department of Biochemistry, St. Mary''s Hospital Medical School, University of London, Norfolk Place, Paddington, London W2 1PG, U.K. Tel. (01) 723 1252 |
| |
Abstract: | We have cloned a 4.9-kb fragment of Pseudomonas aeruginosa DNA containing the structural gene of phospholipase C (PLC), by inserting it into the BamHI site of plasmid pBR322. Strains of Escherichia coli carrying this recombinant plasmid produce PLC, but expression of the gene differs from that in P. aeruginosa in two respects: (i) synthesis of the enzyme appears to be constitutive, i.e., not repressible by the presence of inorganic phosphate in the growth medium, and (ii) most of the enzyme remains associated with the outer membrane instead of being secreted. Insertion mutagenesis at a unique restriction site within the PLC gene destroyed the ability of the plasmid to code, in maxicells, for phospholipase C activity and for an Mr 80000 polypeptide. |
| |
Keywords: | Recombinant DNA constitutive expression plasmid pBR322 vector maxicells insertion mutagenesis outer membrane association enzyme secretion Ap ampicillm kb kilobase pairs PLC phospholipase C SDS sodium dodecyl sulfate Tet tetracycline [ ] indicates plasmid-carrier state 4 deletion |
本文献已被 ScienceDirect 等数据库收录! |
|