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Prokaryotic expression and purification of human hepatic stimulator substance
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 1
- DOI:
- 10.3321/j.issn:0371-0874.2002.01.008
- Key Word:
- 人肝刺激因子;融合蛋白;原核表达
Abstract： 利用基因重组技术,构建成人肝刺激因子(hHSS)和谷胱甘肽转移酶(GST)融合表达载体,转化大肠杆菌BL-21 (DE3),以His·Tag亲和层析纯化表达产物,Factor Xa切割分离hHSS单体,并检测其生物学活性.结果显示,在pET-42a表达体系中hHSS以可溶性蛋白和包涵体两种形式存在,GST-hHSS表达量占菌体可溶性蛋白的30%;Factor Xa切割GST与hHSS之间肽腱,得到33 和15 kD两条蛋白带,经Western杂交证实33 kD条带为GST,而15 kD条带的分子量与hHSS基因序列推测蛋白结果相符.经His·Tag再次纯化可获得hHSS单体,初步证实重组hHSS具有促进肝癌细胞增殖活性.
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