| 小鼠Tnfaipl基因的克隆、表达及抗体制备 |
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| 引用本文： | 刘鑫,韩梅.小鼠Tnfaipl基因的克隆、表达及抗体制备[J].中华医学研究杂志,2005,5(12):1238-1240,F0003. |
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| 作者姓名： | 刘鑫 韩梅 |
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| 作者单位： | [1]湖南师范大学生命科学学院基因敲除与转基因动物研究室,湖南长沙410081 [2]南华大学生命科学与技术学院,湖南衡阳421001 |
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| 摘 要： | 目的克隆小鼠Tnfaipl基因，并在大肠杆菌中表达，通过免疫新西兰兔，获得小鼠Tnfaipl多克隆抗体，为进一步研究小鼠Tnfaipl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Tnfaipl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-Tnfaipl，并用其免疫新西兰兔，获得小鼠Tnfaipl多克隆抗血清。结果得到了小鼠Tnfaipl多克隆抗体。结论成功得到了特异性好的、效价高的小鼠Tnfaipl多克隆抗体。
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| 关 键 词： | 克隆 Tnfaipl 表达 多克隆抗体 |
| 文章编号： | 1680-6115（2005）12-1238-03 |
| 收稿时间： | 2005-04-26 |
Cloning and expression of mouse Tnfaipl and preparation of its polyclonal antibody |
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| LIU Xin, HAN Mei.Cloning and expression of mouse Tnfaipl and preparation of its polyclonal antibody[J].JOurnal of Chinese Medicine Research,2005,5(12):1238-1240,F0003. |
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| Authors: | LIU Xin HAN Mei |
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| Abstract: | Objective Cloning mouse Tnfaipl 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 Tnfaipl. Methods The Tnfaipl cDNA was amplified from mouse liver tissue by RT-PCR, and cloncd into pMD18-T vector. Correct clone was confinned by DNA sequencing. It was then subcloned into expression vector pGEX-4T-2 to express fusion protein GST - Tnfaipl in E. coli. and the purified fusion protein was used to immunize New Zealand rabbits. Results Polyclonal antibody against Tnfaipl was obtained by immunizing the rabbits. Conclusion High levels of specific polyclonal antibody against Tnfaipl was obtained successfully. |
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| Keywords: | clone Tnfaipl expression polyclonal antibody |
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