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A new Escherichia coli strain producing human tumor necrosis factor

机译:产生人肿瘤坏死因子的新大肠杆菌菌株

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摘要

An Escherichia coli strain producing human tumor necrosis factor (TNF-alpha) was obtained using a semisynthetic gene partially optimized in respect of codon composition and a phage T7 promoter. The expression product was accumulated in cells as inclusion bodies in a yield of 50-70 mg/l of culture medium. The recombinant TNF-alpha in the form of inclusion bodies was used for immunization of rats to give a polyclonal antiserum. The resulting antibodies were specific under the immunoblotting conditions to the antigen used for the immunization. A dilution-based refolding procedure was developed; it provided a yield of soluble protein exceeding 85%.
机译:使用在密码子组成和噬菌体T7启动子方面部分优化的半合成基因,获得了产生人肿瘤坏死因子(TNF-α)的大肠杆菌菌株。表达产物以50-70mg / l的培养基产量积累在细胞中作为包涵体。以包涵体形式的重组TNF-α用于大鼠的免疫,得到多克隆抗血清。所得抗体在免疫印迹条件下对用于免疫的抗原具有特异性。开发了基于稀释的重折叠程序;它提供了超过85%的可溶性蛋白质收率。

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