The Construction, Expression and Purification of Therapeutic Adjuvant-free Protein Vaccine——HPV16z-Hsp65-E6/E7
- VernacularTitle:HPV16~+治疗性无佐剂蛋白疫苗——HPV16z-Hsp65-E6/E7的构建、表达及纯化工艺研究
- Author:
Xiaobing WANG
;
Mo LI
;
Yi LIU
;
Haimei TIAN
;
Zhaoyang LIU
;
Yanfen LI
;
Dongyan CAO
;
Zhi LIANG
;
Dongwan CHENG
;
Changjun SHAO
;
Wei ZHANG
;
- Publication Type:Journal Article
- Keywords:
HPV16 Therapeutic vaccine Cervical cancer
- From:
China Biotechnology
2006;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.
