The complete nucleotide sequence and immunochemistry of the major outer membrane protein of Neisseria gonorrheae

Butt, Neil James (1991) The complete nucleotide sequence and immunochemistry of the major outer membrane protein of Neisseria gonorrheae. University of Southampton, Doctoral Thesis.

Abstract

The major outer membrane protein of Neisseria gonorrhoeae has been found to elicit an immune response in both localised and invasive disease. The response has been shown to generate limited immunity to further infection. In this study the PIB protein from Neisseria gonorrhoeae strain P9 was cloned and sequenced using a combination of restriction endonuclease fragments and the Polymerase chain reaction. This gave a sequence highly homologous to other PI sequences with variation occurring in only two regions. A series of synthetic peptides were produced corresponding to the derived protein sequence. These were reacted with monoclonal antibodies to try and determine any specific epitopes within the protein. The epitopes of 5 antibodies were found of which 4 were type specific and one was cross reactive with 92% of all gonococcal PIB strains tested. To investigate the type specific reactivity further the polymerase chain reaction was used to amplify the variable regions of 6 other strains which were known to react with one or more of the antibodies tested. The resulting sequences confirmed that only two variable regions existed. On reactivity with peptides made to these strains it appeared that only one of these regions reacted in this study. The epitopes for 3 type specific monoclonal antibodies were discovered by reacting the antibodies with these peptides. Using the sequence obtained in this study and data from other Neisserial porins a topological model of the P9 PIB was constructed, showing why certain regions were more immunogenic than others in respect to protein surface exposure.

This record has no associated files available for download.

Contributors

Download statistics