Stable expression of meningococcal class 1 protein in an antigenically reactive form in outer membranes of Escherichia coli

Molecular Microbiology
D A WhiteJ E Heckels

Abstract

The entire gene encoding the class 1 outer membrane protein of Neisseria meningitidis is located on a 2.2kb fragment, obtained on digestion of chromosomal DNA with Xbal. This Xbal fragment from strain MC50 (subtype P1-16), which had previously been cloned in bacteriophage M13, has been transferred to the plasmid vector pMTL20. The resulting plasmid (pPORA100) was propagated in Escherichia coli (JM109) and cell lysates were subjected to SDS-PAGE. Western blotting with anti-class 1 protein antibodies revealed constitutive expression of a protein of 41 kD, corresponding to the class 1 protein of the parent meningococcal strain, which was absent in the E. coli control. Fractionation of E. coli cells carrying the recombinant plasmid revealed that the protein was exclusively located in the outer membrane, and N-terminal amino acid analysis of the expressed protein revealed that normal processing of the signal peptide had occurred. Immuno-gold electron microscopy showed that the protective epitope recognized by a P1-16 subtype-specific monoclonal antibody was exposed in an antigenically reactive form on the surface of E. coli cells carrying plasmid pPORA100. In contrast, expression in E. coli of a second plasmid (pPORA104) lacking the...Continue Reading

References

Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Apr 1, 1989·Clinical Microbiology Reviews·S E West, V L Clark
Apr 1, 1989·Clinical Microbiology Reviews·P J Hitchcock
Nov 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·E C GotschlichM Koomey
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·N H Carbonetti, P F Sparling
Dec 1, 1988·FEMS Microbiology Immunology·H Abdillahi, J T Poolman
Jun 1, 1985·The Journal of Hand Surgery : Journal of the British Society for Surgery of the Hand·N J Treble
Aug 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·S N CohenL Hsu
Jan 1, 1974·Methods in Enzymology·M J Osborn, R Munson
Jun 1, 1969·The Journal of Experimental Medicine·I GoldschneiderM S Artenstein
Jul 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·D DiMaioT Maniatis
Feb 1, 1981·Journal of Bacteriology·J E Heckels

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Citations

May 18, 2007·Annual Review of Microbiology·Martine P BosJan Tommassen
Sep 1, 1991·Infection and Immunity·P van der LeyJ T Poolman

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