Elastase gene expression in non-elastase-producing Pseudomonas aeruginosa strains using novel shuttle vector systems.
In order to determine whether non-elastase-producing strains of Pseudomonas aeruginosa such as N-10, PA103 and IFO3080 can express foreign elastase genes, we introduced elastase genes from P. aeruginosa IFO3455 (elastase-producing) as well as from PA103 and N-10 into non-elastase-producing P. aeruginosa strains. Results suggested that gene expression, secretion, and precursor processing systems of elastase were essentially normal in P. aeruginosa N-10 and IFO3080. Our studies using various elastase genes showed that both the elastase structural gene and 5'-upstream regions of P. aeruginosa PA103 were also normal. This was confirmed by the finding that P. aeruginosa N-10 and IFO3080 which carry the PA103 elastase gene produced elastase. Several deleted or chimeric genes were constructed using the 5'-upstream regions of elastase genes from P. aeruginosa N-10 or PA103 and studies of expression revealed that two individual DNA bases seem to be important in suppressing P. aeruginosa N-10 elastase gene expression. Possible reasons for the lack of elastase expression in these non-elastase-producing strains are discussed.
Ishii
T
T
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
Fukushima
J
J
Fujita
S
S
Shigematsu
T
T
Ando
N
N
Ishiwata
T
T
Kurata
M
M
Kawamoto
S
S
Morihara
K
K
Okuda
K
K
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000096:biosynthesis* / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012333:RNA, Messenger; D010196:Pancreatic Elastase