Expression and purification of Litopenaeus vannamei allergen protein Lit v1.2
10.3969/j.issn.1000-484X.2015.12.016
- VernacularTitle:凡纳滨对虾过敏原蛋白Lit v 1.2的原核表达与纯化
- Author:
Huifang CHEN
;
He LAI
;
Yuyi HUANG
;
Zehong ZOU
;
Ying HE
;
Ailin TAO
;
Wen LI
- Publication Type:Journal Article
- Keywords:
Litopenaeus vannamei;
Allergen;
Tropomyosin;
Recombinant expression;
Protein purification
- From:
Chinese Journal of Immunology
2015;31(12):1659-1662
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To obtain purified recombinant Litopenaeus vannamei allergen protein Lit v 1.2.Methods: The target gene of Lit v 1.2 was inserted into clone vector pGEM-T and then ligated to the expression vector pET 44a.The pET44a-Liv 1.2 was transformed into Rosetta and screened by ampicillin resistance .The recombinant protein was expressed by IPTG induction .The protein was purified by 6-His tag affinity chromatography and the purification was analyzed by SDS-PAGE gel electrophoresis .Results:The ex-pression plasmid pET44a-Lit v 1.2 was constructed.SDS-PAGE showed that expressed Lit v 1.2 was efficient and soluble in E.coli Rosetta.The protein molecular weight was consistent with the theoretical value .The highly purified target protein was obtained.Conclusion:In this study ,we successfully gained highly purified recombinant allergen protein Lit v 1.2 which was expressed in prokaryotic system and purified by affinity chromatography column .The purified Lit v 1.2 protein will facilitate us to further study its role in immunological responses .
