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为了提高人肿瘤坏死因子α(hTNF-α)靶向性,同时降低其在体内的毒副作用,应用现代基因工程技术对hTNF-α进行改造,构建融合蛋白成为新的途径。本文利用基因工程的方法成功构建hTNF-α与基质金属蛋白酶-1(MMP1)f、oldon序列的重组质粒,将该质粒转化至Rosetta2(DE3),在一定条件下,经异丙基--βD-硫代半乳糖苷(IPTG)诱导融合蛋白可溶性表达,后用谷胱甘肽(GST)树脂纯化试剂盒进行纯化,得到融合蛋白的纯化产物。为以后的科研和临床应用打下基础。
In order to improve the targeting of human tumor necrosis factor α (hTNF-α) and reduce its toxicity in vivo, it is a new way to construct hTNF-α by using modern genetic engineering technology. In this paper, the recombinant plasmid of hTNF-αand MMP-1f, oldon sequences was successfully constructed by genetic engineering and transformed into Rosetta2 (DE3). Under certain conditions, (IPTG) induced fusion protein was purified and purified by GST resin purification kit to obtain the purified product of the fusion protein. Lay the foundation for future scientific research and clinical application.