【摘要】 目的 克隆小鼠Tnfaip1基因，并在大肠杆菌中表达，通过免疫新西兰兔，获得小鼠Tnfaip1多克隆抗体，为进一步研究小鼠Tnfaip1的功能奠定基础。方法 用RT-PCR方法从小鼠肝脏组织中扩增获得Tnfaip1基因的蛋白质编码区，将目的片段连接到PMD18-T载体，然后亚克隆至pGEX-4T-2中，进而在大肠杆菌BL21(DE3)中表达融合蛋白质GST- Tnfaip1，并用其免疫新西兰兔，获得小鼠Tnfaip1多克隆抗血清。结果 得到了小鼠Tnfaip1多克隆抗体。结论 成功得到了特异性好的、效价高的小鼠Tnfaip1多克隆抗体。

    【关键词】 克隆；Tnfaip1；表达；多克隆抗体

    Cloning and expression of mouse Tnfaip1 and preparation of its polyclonal antibody

    LIU Xin，HAN Mei.

    Laboratory of Gene Knock-out and Transgenic Animal，College of Life Science，Hunan Normal University，Changsha，Hunan 410081，China

    【Abstract】 Objective Cloning mouse Tnfaip1 and expressing its protein in E.coli and preparing the polyclonal antibody by immunizing the rabbit to pave the way for understanding the role of Tnfaip1.Methods The Tnfaip1 cDNA was amplified from mouse liver tissue by RT-PCR，and cloned into pMD18-T vector. Correct clone was confirmed by DNA sequencing. It was then subcloned into expression vector pGEX-4T-2 to express fusion protein GST -Tnfaip1 in E.coli. and the purified fusion protein was used to immunize New Zealand rabbits.Results Polyclonal antibody against Tnfaip1 was obtained by immunizing the rabbits.Conclusion High levels of specific polyclonal antibody against Tnfaip1 was obtained successfully. ......