Cloing and High Level Expression of VP6 Gene From Group A Human Rotavirus in E.coli
- VernacularTitle:A组人轮状病毒VP6基因克隆及在大肠杆菌中的高效表达
- Author:
Zhi-Liang CAO
;
Yu-Ling WEN
;
Yuang-Ding CHEN
;
- Publication Type:Journal Article
- Keywords:
Rotavirus VP6 Prokaryotic expression
- From:
China Biotechnology
2006;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
The structural protein VP6 of rotavirus form the middle layer of the triple-layered viral capsid, playing a key role in the organization of the virion. The gene of structural protein 6 of rotavirus strain TB-Chen isolated from a clinic sample was amplified using PCR from the reverse transcription product of RV genome RNA, using pET as expression vector, a recombinant plasmid pET-VP6 containing coding sequence of VP6 protein was constructed. The results showed that the VP6 was highly efficiently expressed in E. coli BL21(DE3) cells which were transformed with the recombinant plasmid pET-VP6.The expressed VP6 protein possessed 27.4% of total cells protein, with an approximately 45kDa of molecular weight, and could be recognized by guinea pig anti-SA11 antibody on Western blot. The results obtained provide important basis for further study on structure and function of the VP6 protein.
