Cloning and characterization of elastase genes from Pseudomonas aeruginosa.
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.
D000942:Antigens, Bacterial / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D005786:Gene Expression Regulation; D005796:Genes; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D010196:Pancreatic Elastase / Q000235:genetics* / Q000276:immunology; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics*
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D010196:Pancreatic Elastase