Construction of a recombinant Neisseria gonorrhoeae strain with rmp gene deletion and study on the bactericidal activities of its antibodies
10.3760/cma.j.issn.0254-5101.2013.08.004
- VernacularTitle:淋病奈瑟菌rmp基因缺失突变株的构建及其抗体杀菌作用研究
- Author:
Guocai LI
;
Rushan XIE
;
Yanli MAO
;
Shuangxi LIU
;
Hongmei JIAO
;
Hongju CHEN
;
Hua YAN
;
Mingchun JI
- Publication Type:Journal Article
- Keywords:
Neisseria gonorrhoeae;
Homologous recombination;
rmp gene deletion mutants;
Live attenuated vaccine
- From:Chinese Journal of Microbiology and Immunology
2013;(8):578-582
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.
