中国免疫学杂志2015，Vol.31Issue(12)：1659-1662,4.DOI:10.3969/j.issn.1000-484X.2015.12.016
凡纳滨对虾过敏原蛋白Lit v 1.2的原核表达与纯化
Expression and purification of Litopenaeus vannamei allergen protein Lit v1.2
摘要
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 .关键词
凡纳滨对虾/过敏原/原肌球蛋白/重组表达/蛋白纯化Key words
Litopenaeus vannamei/Allergen/Tropomyosin/Recombinant expression/Protein purification分类
医药卫生引用本文复制引用
陈惠芳,赖荷,黄于艺,邹泽红,何颖,陶爱林,李文..凡纳滨对虾过敏原蛋白Lit v 1.2的原核表达与纯化[J].中国免疫学杂志,2015,31(12):1659-1662,4.基金项目
本文受国家重大科技专项(2014ZX08011-005B)、广东高校科技创新项目(2013KJCX148)和广州市科技攻关(201300000159)项目的资助. （2014ZX08011-005B）
