The Procaryotic Expression, Purification and Activity Analysis of VIP-sTNFRII
- VernacularTitle:sTNFRⅡ和VIP融合蛋白的基因克隆表达、纯化及活性检测
- Author:
Hong WANG
;
Wei-Sen ZENG
;
Jin-Hua CHEN
;
Shan-Shan WANG
;
Dan LIU
;
Yan-Ni YANG
;
Bai-Hong CHEN
;
Ling LI
;
- Publication Type:Journal Article
- Keywords:
sTNFRII VIP Fusion protein Expression Purification
- From:
China Biotechnology
2006;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
A prokaryotic expression plasmid containing VIP (vasoactive intestinal peptide) and sTNFRII(soluble tumor necrosis factor receptor II ) genes was constructed. The sTNFRII was cloned by PCR by using special primers which contained VIP gene ORF and a linker in its forward primer. The amplified fragment was inserted into the expression vector pET32a between BamHI and Hind III restriction sites. Transformed E.coli DH5 by pET32a-VIP- sTNFRIIexpressed the fusion protein. After being identified, the protein was purified by ion exchange chromatography and by hydrophobic interaction chromatography. The reconstructed protein showed high bio-activity and could be applied for further use.
