Cloning and expression in Escherichia coli of opc, the gene for an unusual class 5 outer membrane protein from Neisseria meningitidis (meningococci/surface antigen).
A genomic library was constructed in a lambda gt11 vector using chromosomal DNA from a meningococcal serogroup A strain and plaques expressing the class 5C protein were recognized by screening with specific monoclonal antibodies. The opc insert was subcloned into a multicopy plasmid which induced expression of that protein in Escherichia coli as a surface-exposed major outer membrane protein. The nucleotide sequence of opc is typical of an outer membrane protein with a promoter and terminator region, a leader peptide which is cleaved during expression and a complete open reading frame. Unlike other meningococcal class 5 proteins or gonococcal P.II proteins, the sequence did not contain any pentanucleotide repeats and the sequence showed little homology to these other functionally related proteins. However, the predicted amino acid sequence of the mature protein for opc showed 27% similarity to that for a second opa gene cloned from the same meningococcal strain. This is the first report of cloning and expression of a functional meningococcal gene encoding a class 5 outer membrane protein in E. coli.
Olyhoek
A J
AJ
Max-Planck Institut für molekulare Genetik, Berlin, Germany.
Sarkari
J
J
Bopp
M
M
Morelli
G
G
Achtman
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D014176:Protein Biosynthesis; D012091:Repetitive Sequences, Nucleic Acid
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C074517:opc protein, bacteria