Open AccessCloning and characterization of elastase genes from Pseudomonas aeruginosa
Author(s) -
Péter Schád,
Robert Bever,
Thalia I. Nicas,
Frédéric Leduc,
Larry F. Hanne,
Barbara H. Iglewski
Publication year - 1987
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.169.6.2691-2696.1987
Subject(s) - biology , ecori , elastase , pancreatic elastase , plasmid , escherichia coli , pseudomonas aeruginosa , microbiology and biotechnology , molecular cloning , restriction map , restriction enzyme , structural gene , pseudomonadaceae , gene , biochemistry , bacteria , enzyme , genetics , peptide sequence
A gene bank was constructed from Pseudomonas aeruginosa PAO1 and used to complement three P. aeruginosa elastase-deficient strains. One clone, pRF1, contained a gene which restored elastase production in two P. aeruginosa isolates deficient in elastase production (PA-E15 and PAO-E105). This gene also encoded production of elastase antigen and activity in Escherichia coli and is the structural gene for Pseudomonas elastase. A second clone, pHN13, contained a 20-kilobase (kb) EcoRI insert which was not related to the 8-kb EcoRI insert of pRF1 as determined by restriction analysis and DNA hybridization. A 2.2-kb SalI-HindIII fragment from pHN3 was subcloned into pUC18, forming pRB1822-1. Plasmid pRB1822-1 restored normal elastolytic activity to PAO-E64, a mutant for elastase activity. Clones derived from pHN13 failed to elicit elastase antigen or enzymatic activity in E. coli.