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Expression of the serratia marcescens lipo protein gene in escherichia coli


, : Expression of the serratia marcescens lipo protein gene in escherichia coli. Journal of Bacteriology 146(3): 861-866

The lipoprotein gene (lpp) of S. marcescens was cloned in a .lambda. phage vector. This lpp gene was recloned in plasmid vectors pBR322 and pSC101. When a lipoprotein-deficient (lpp) mutant of E. coli was transformed with pBR322 carrying the S. marcescens lpp gene, cells became nonleaky for RNase, resistant to EDTA and sensitive to globomycin. The lipoprotein was found exclusively in the outer membrane fraction. The S. marcescens lipoprotein was normally secreted across the cytoplasmic membrane, modified and assembled in the E. coli outer membrane. The amount of the free-form lipoprotein produced in this system was 3 times higher than that produced in lpp+ E. coli cells; there was no difference in the amount of the bound-form lipoprotein. lpp E. coli cells which harbored pSC101 carrying the S. marcescens lpp gene produced only 1/3 of the free-form lipoprotein produced in lpp E. coli cells which harbored pSC101 carrying the E. coli lpp gene. One major factor causing this difference in efficiency of gene expression between the lpp genes of S. marcescens and E. coli appears to be a deletion mutation at the transcription termination region found in the cloned S. marcescens lpp gene. The functional half-life of the S. marcescens lpp mRNA in E. coli was about half that of the E. coli lpp mRNA.

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