中国免疫学杂志Issue(12)：1666-1669,4.DOI:10.3969/j.issn.1000-484X.2014.12.17
HE4两种融合蛋白的基因构建和原核表达
Construction and expression of two kinds of HE4 fusion protein
张雪峰 1刘一兵 2李子颖 1贾娟娟 2王斌 1韩世泉2
作者信息
- 1. 原子高科股份有限公司，北京 102413
- 2. 中国原子能科学研究院，北京102413
- 折叠
摘要
Abstract
Objective:To express human epididymis protein 4 (HE4) in prokaryotic cells,purify the expressed product and de-termine its activity by immunoassay kit.Methods: The gene encoding HE 4 was cloned using RT-PCR technique from total RNA of ovarian carcinoma cells ES-2,the amplified HE4 gene was cloned into prokaryotic expression vector pGEX-4T-1 and PET26b respective-ly.The recombinant plasmid pGEX-4T-1-HE4 and PET26b-HE4 were constructed and transformed into E.coli BL21 cells respectively, and protein ( GST-HE4 and His-HE4 ) expressions were induced by IPTG and identified by SDS-PAGE and commercial ELISA kit.Results:Restriction analysis and sequencing proved that recombinant plasmid pGEX -4T-1-HE4 and PET26b-HE4 were constructed correctly.The expressed recombinant proteins ,with the relative molecular mass of about 38 000 and 12 000 ,showed specific binding to monoclonal antibody against HE 4.Conclusion:Two kinds of recombinant HE 4 protein are successfully expressed in prokaryotic cells , which laid a foundation of preparation of immunoassay reagents.关键词
卵巢癌/人附睾蛋白4 ( HE4 )/克隆/表达Key words
Ovarian carcinoma/Human epididymis protein 4( HE4)/Cloning/Expression分类
医药卫生引用本文复制引用
张雪峰,刘一兵,李子颖,贾娟娟,王斌,韩世泉..HE4两种融合蛋白的基因构建和原核表达[J].中国免疫学杂志,2014,(12):1666-1669,4.
