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将人γ干扰素/β肿瘤坏死因子融合蛋白(hIFN-γ/TNF-β,rhTNF-β)是一种颇具理论研究价值和临床应用前景的新型细胞因子.鉴于现有的rhTNF-β表达质粒的表达水平较低(1%左右),不能满足研究工作的需要.本工作在过去构建rhTNF-β表达质粒的基础上,采用分步构建的方式,首先构建了该融合蛋白基因5’端部分的人γ干扰素突变体(hIFN-γ134-X13)编码序列的表达质粒,在获得高效表达以后,再在其3’端连接上5’端缺失23个氨基酸残基(aa)的人β肿瘤坏死因子(hTNF-β148)的核酸序列,进行整个融合蛋白基因的表达研究.实验结果表明,新构建的rhTNF-β表达质粒经转化大肠杆菌BL21(DE3),在λP_RP_L启动子的控制下,温敏诱导表
It is a new type of cytokines with theoretical research value and clinical application prospect.Human interferon / β tumor necrosis factor fusion protein (hIFN-γ / TNF-β, rhTNF-β) (1%), which can not meet the needs of research work.In this work, based on the construction of rhTNF-β expression plasmid in the past, a 5-terminal part of the fusion protein gene (HIFN-γ134-X13) coding sequence of the human interferon-γ (hIFN-γ134-X13) coding sequence was obtained, and after high-level expression, a human β-tumor with a deletion of 23 amino acid residues (HTNF-β148), the expression of the fusion protein gene was studied.The experimental results showed that the newly constructed rhTNF-β expression plasmid was transformed into E. coli BL21 (DE3) under the control of λP_RP_L promoter Sensitive induction table