Cloning and expression of a gene coding for the prolipoprotein signal peptidase of Escherichia coli.
An Escherichia coli mutant, Y815, has a temperature-sensitive prolipoprotein signal peptidase. IPTG-induced synthesis of the major outer membrane prolipoprotein (PLP) results in the inhibition of cell growth because of accumulation of PLP in its envelope [J. Bacteriol. (1982) 152, 1163-1168]. The 2000 E. coli strains of Clarke and Carbon's collection were screened for the presence of a plasmid complementing the IPTG-sensitivity of the growth of Y815. One plasmid, pLC3-13, complemented the IPTG-sensitivity. The envelope fraction prepared from Y815 transformed by pLC3-13 showed high activity of the PLP signal peptidase in vitro at high temperature. A 4 kb AccI fragment subcloned onto plasmid pHY001 was shown to carry the gene for the PLP signal peptidase.
D002874:Chromosome Mapping; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005815:Genetic Code; D008565:Membrane Proteins; D009154:Mutation; D010957:Plasmids; D012697:Serine Endopeptidases
D008565:Membrane Proteins; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase