Preparation and Identification of Recombinant Vasoactive Intestinal Peptide
- VernacularTitle:重组血管活性肠肽的制备及鉴定
- Author:
Shan-Shan XIE
;
Rong-Jie YU
;
Le ZENG
;
Juan LI
;
Jing-Jing WANG
;
An HONG
;
- Publication Type:Journal Article
- Keywords:
Vasoactive intestinal peptide Intein Purification and identification Anti-inflammatory effects
- From:
China Biotechnology
2006;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
In order to prepare the recombinant vasoactive intestinal peptide (VIP) using intein mediated rapid purification system,the cDNA encoding the recombinant VIP was designed and synthesized according to the preference of E.coli,and then was cloned into the expression vector PTWIN. The recombinant plasmid PTWIN-VIP was transformed into expression host E.coli strain ER2566.The fusion protein consisting of the recombinant VIP,intein and chitin binding domain was expressed and purified by chitin affinity chromatography. The target peptide was released from the fusion protein by changing the temperature and the pH of the cleavage buffer. The molecular weight of the recombinant VIP was determined by the mass spectrometry and the results was conformity with the theoretical value. The preliminary bioactivity assay indicated that the recombinant VIP decreased the serum resistin levels significantly in LPS-induced acute inflammation. The preparation and the characterization of anti-inflammatory effects of the recombinant VIP layed the foundation for its further application.
