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重组人PD-1IgV融合蛋白的克隆、表达、纯化及生物学活性检测 被引量：1

Cloning,expression,purification and biologic characteristics of rhPD-1IgV protein
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摘要 目的:获得具有生物学活性的重组人PD-1IgV(rh-PD-1IgV)蛋白.方法:人工合成PD-1IgV基因,并将该基因克隆入原核表达载体pQE-30中.转化宿主细胞大肠杆菌DH5α,经IPTG诱导表达重组融合rhPD-1IgV蛋白.对表达产物进行SDS-PAGE电泳和Westernblot检测分析.结果:rhPD-1IgV的Mr约为15×103,与根据其氨基酸组成计算的理论值一致.成功纯化了rhPD-1IgV蛋白,并对其进行了复性和浓缩,且复性后的蛋白可与其配体B7-H1结合,显示出良好的结合活性.结论:成功地克隆、表达和纯化了rhPD-1IgV蛋白. AIM： To obtain biologically active recombinant human PD-1IgV （rhPD-1IgV） protein. METHODS： Gene encoding for rhPD-1IgV was synthesized and cloned into the expression vector pQE-30. The expression plasmid was then trans- formed into E. coli DH5α. The expression of the fusion protein rhPD-1IgV was induced with IPTG and detected by SDS-PAGE and Western blot. The biological activity of rhPD-1IgV was identified by ELISA and flow cytometry. RESULTS： SDS-PAGE showed that the molecular weight of rhPD-1IgV was about 15 × 10^3, which was consistent with the theoretical value calculated based on the composition of amino acid. Purified rhPD-1IgV was refolded, concentrated and associated well with BT-H1. CONCLUSION： rhPD-1IgV fusion protein is successfully cloned, expressed and purified. The protein associated with B7-H1 has a strong binding activity.
作者 王伟华 张英起 颜真
机构地区 第四军医大学药学系生物技术中心
出处 《第四军医大学学报》 北大核心 2006年第5期408-411,共4页 Journal of the Fourth Military Medical University
基金 教育部"长江学者和创新团队发展计划"(IRT0459)
关键词 PD-1IgV 重组融合蛋白 纯化 复性 pQE-30载体 PD-1IgV recombinant fusion protein purification refolding pQE-30 vector
分类号 Q78 [生物学—分子生物学]
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参考文献10

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共引文献1

同被引文献10

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引证文献1

第四军医大学学报
2006年 第5期

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