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%.
出处
《华东理工大学学报（自然科学版）》
CAS
CSCD
北大核心
2004年第4期465-469,共5页
Journal of East China University of Science and Technology
基金
上海-SK研究与发展基金资助项目(2002002-S)。
关键词
3A蛋白
包涵体
复性
分离纯化
protein 3A
inclusion body
refolding
purification
