重组人canstatin表达产物的抗肿瘤活性鉴定
Identification of Recombinant Human Canstatin s Biological Activity
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摘要: 目的 正确构建人canstatin原核表达载体, 诱导表达重组人canstatin融合蛋白, 并对其活性鉴定。方法 将人canstatincDNA亚克隆入 pQE30, 构建重组质粒 pQE30 canstatin ;转化M 15 pREP4 中, IPTG诱导表达, 纯化回收表达产物, 复性后用Lewis肺癌移植瘤模型对其活性鉴定。结果 成功构建人canstatincDNA表达载体, 纯化回收, 获得高纯度canstatin重组蛋白。纯化产物在体内能抑制Lewis肺癌移植瘤血管的生成, 从而抑制肿瘤生长和转移, 抑瘤率 (% )为 6 0 .0 %, 转移抑制率 (% )为 74 .3%。结论 (1)成功构建了原核表达载体 pQE30 canstatin。 (2 )重组人canstatin融合蛋白在原核表达系统中高水平表达, 并获得高纯度canstatin重组蛋白。 (3)重组canstatin融合蛋白可抑制Lewis肺癌移植瘤血管的生成, 从而抑制肿瘤生长和转移活性。Abstract: Objective To construct human canstatin gene prokaryotic expression vector. To express recombinant human canstatin in E.coli M15_ and investigate its biological activity. Methods The obtained canstatin cDNA fragment was subcloned into the BamHⅠand HindⅢsites of pQE30 prokaryotic expression vector. The recombinant protein was expressed with induction of IPTG in E.coli M15_ . After the purification renaturation of the expression product,its anti-angiogenesis biological activity ws detected by syngeneic m...
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