Immunoreactivity analysis of Tp47 recombinant protein of Treponema Pallidum by Western-Blot assay
10.3969/j.issn.1673-4130.2015.10.004
- VernacularTitle:免疫印迹法测定Tp47蛋白与梅毒患者血清的免疫反应性
- Author:
Heping ZHENG
;
Xiaolin QIN
;
Jinmei HUANG
;
Yaohua XUE
;
Shun BAI
;
Ping LV
;
Bin YANG
- Publication Type:Journal Article
- Keywords:
Treponema pallidum;
recombinant protein;
Western-Blot
- From:
International Journal of Laboratory Medicine
2015;(10):1327-1329
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone ,construction ,express and purify Tp47 of Treponema pallidum (Tp) ,and assess the immunoreac‐tivity by Western‐Blot .Methods Tp47 was amplified by polymerase chain reaction ,and then cloned to the vector pGEX‐6P‐1 .The correct sequence of the recombinant plasmids pGEX‐6P1‐Tp47 was transformed into Escherichia coli BL21 (DE3) and induced .The expression product was analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western‐Blot .The expression protein was purified .Serum of different clinical stages of syphilis was used as the antibody to detect the immunoreactivity of the protein by Western‐Blot .Results A fusion protein with molecular weight about 71 × 103 was attained .Western‐Blot proved that the recombinant protein can react with Tp IgG positive sera .And the specificities and sensitivities of the diagnostic reagent detected by sera were 100% .Conclusion The recombinant protein Tp47 was expressed and purified with good antigen activity ,which could provide the basis of theory and practice for the development of early diagnostic kit applying to detect Tp infection .
