【摘要】 目的:肿瘤坏死因子(Tumor necrosis factor,TNF)是一种具有广泛生物学效应的细胞因子,在机体炎症和免疫防御反应中起着重要作用,其中包括运动刺激所产生的机体反应。一种新型蛋白TTRAP被认为在TNF信号通路中有可能起重要作用,为揭示其作用以及寻找与其作用的互作蛋白,构建了TTRAP酵母双杂交系统的"诱饵"载体。方法:对含有人TTRAP全部序列的表达载体pGEX-4T-1-TTRAP进行PCR扩增,酶切后将TTRAP基因连接到酵母双杂交系统的pLexA质粒上,得到"诱饵"(Bait)-pLexA-TTRAP,导入大肠杆菌扩增,筛选得到阳性克隆并提取重组质粒。结果:测序结果表明克隆正确。结论:成功构建pLexA-TTRAP酵母双杂交载体,为寻找TTRAP的互作蛋白打下基础。更多还原

【Abstract】 Tumor necrosis factor(TNF)is a cytokine with wide biological effect and plays the important roles in anti-inflammatory and immune responses including exercise stress.A novel protein TTRAP has been identified which may be important in the TNF transduction pathway. In order to reveal its function and screen the interactive protein with TTRAP,the bait in yeast two-hybrid system of TTRAP was constructed.Amplifying the full length human TTRAP gene in the vector of pGEX-4T-1-TTRAP then the product was cloned into the plasmid of pL- exA in yeast two-hybrid system after digested with endonucleases.The(Bait)-pLexA-TTRAP was obtained and transformed into E.coli.The positive clones were screened for and the re- combined bait plasmid had been distilled.The sequence analysis indicated that the done was correct.It concluded that the vector of pLexA-TTRAP had been constructed successfully which would provide a foundation for seeking the interactive protein with TTRAP.更多还原