The expression and preliminary evaluation of HPV6bL2deltaN360E7E6 fusion protein in E. coli for genital warts.
- Author:
Zheng PANG
1
;
Li ZHAO
;
Jiao REN
;
Jing FENG
;
Zhong-Xian ZHANG
;
Wen-Jie TAN
;
Li RUAN
;
Hou-Wen TIAN
Author Information
- Publication Type:Journal Article
- MeSH: Adjuvants, Immunologic; Animals; Condylomata Acuminata; genetics; Enzyme-Linked Immunosorbent Assay; Enzyme-Linked Immunospot Assay; methods; Escherichia coli; genetics; Gene Expression; Genetic Vectors; Immunity, Humoral; Immunization; Mice; Mice, Inbred C57BL; Polymerase Chain Reaction; Recombinant Fusion Proteins; genetics; metabolism; pharmacology; Recombinant Proteins; chemistry; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2010;24(3):171-174
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo express HPV6bL2deltaN360E7E6 fusion protein in E. coli and preliminarily evaluate its immune effect.
METHODSThree HPV6b gene fragments, which were L2(1-360 bp), E7 and E6, were fused by overlapping PCR, then were inserted into a prokaryotic expression vector and expressed in E. coli. C57BL/6 mice were immunized with purified fusion protein plus Al (OH)3 and/or CpG adjuvants through intramuscular route, the cellular and humoral immune responses were detected by IFN-gamma ELISPOT and ELISA respectively.
RESULTSProtein plus CpG adjuvant could induce the strongest cellular immune response to E7 and E6, high antibody titer against L2 could be detected in all immunized groups but there were no significant difference among these groups.
CONCLUSIONSHPV6bL2deltaN360E7E6 gene was successfully cloned into pQE30 vector and expressed in E. coli, the fusion protein was also purified and proved that could induce strong cellular and humoral immune responses with appropriate adjuvant in C57BL/ 6 mice and could be used for future research.
