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    尹芳, 陈国豪, 陆兵, 刘叶青, 徐殿胜. 基因工程血管抑素3A蛋白的分离纯化[J]. 华东理工大学学报(自然科学版), 2004, (4): 465-469.
    引用本文: 尹芳, 陈国豪, 陆兵, 刘叶青, 徐殿胜. 基因工程血管抑素3A蛋白的分离纯化[J]. 华东理工大学学报(自然科学版), 2004, (4): 465-469.
    Purification of Recombinant Protein 3A[J]. Journal of East China University of Science and Technology, 2004, (4): 465-469.
    Citation: Purification of Recombinant Protein 3A[J]. Journal of East China University of Science and Technology, 2004, (4): 465-469.

    基因工程血管抑素3A蛋白的分离纯化

    Purification of Recombinant Protein 3A

    • 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Sephadex G75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%。

       

      Abstract: 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%.

       

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