文献类型：期刊文献

中文题名：基因工程血管抑素3A蛋白的分离纯化

英文题名：Purification of Recombinant Protein 3A

作者：尹芳[1];陈国豪[1];陆兵[1];刘叶青[1];徐殿胜[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室生物工程系,上海200237

年份：2004

卷号：30

期号：4

起止页码：465

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

基金：上海-SK研究与发展基金资助项目(2002002-S)。

语种：中文

中文关键词：3A蛋白;包涵体;复性;分离纯化

外文关键词：protein 3A; inclusion body; refolding; purification

摘要：在8mol/mL尿素溶液中,以二硫苏糖醇(DTT)为还原剂,对基因工程血管抑素3A包涵体进行溶解实验。结果发现:1.5h后溶解的上清液蛋白浓度达26.2mg/mL。SDS-PAGE电泳扫描结果显示,3A蛋白经过SephadexG75分离后PAGE电泳纯达到100%,该步收率达85.82%,相对分子质量为10ku。采用分步稀释法对其进行复性,所获复性3A蛋白经MTT法检测表明:3A蛋白浓度为0.1μg/mL时,对内皮细胞的生长抑制率为93.4%。
The inclusion bodies of 3A were dissolved in 8 mol/mL urea buffer, and dithiothreitol(DTT) was added as reducing agent. Result showed that after one and a half hour, the concentration of soluble protein was (26.2) mg/mL.Being separated by sephadex G75 gel filtration chromatography, the recovery and purity of protein 3A determined by SDS-PAGE were 85.82% and 100% respectively, and its molecular weight was 10 ku. By multi-step dilution, the protein 3A was refolded, and the inhibition rates measured by MTT was 93.4%.