| 人睫状神经营养因子的克隆与原核表达 |
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| 引用本文： | 江国春,袁丽珍.人睫状神经营养因子的克隆与原核表达[J].军事医学科学院院刊,1998,22(2):118-121,124. |
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| 作者姓名： | 江国春 袁丽珍 |
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| 作者单位： | 军事医学科学院放射医学研究所 |
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| 摘 要： | 目的：克隆人睫状神经营养因子基因ｃＤＮＡ，并探索其在大肠杆菌中高效表达。方法：提取总ＲＮＡ，逆转录聚合酶链反应（ＲＴ－ＰＣＲ），序列测定正确后原核表达并用制备电泳纯化。结果：克隆了人睫状神经营养因子基因ｃＤＮＡ，实现了其在大肠杆菌中的高效表达，表达量至少达到２６％，制备电泳纯化后，重组蛋白纯度达到９５％。结论：从胎儿坐骨神经组织中克隆到人睫状神经营养因子基因ｃＤＮＡ，制备电泳纯化出高纯度的重组蛋白
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| 关 键 词： | 神经生长因子 聚合酶链反应 基因表达 克隆 CNTF |
Cloning and expression of human ciliary neurotrophic factor cDNA from fetus |
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| Jiang Guochun,Yuan Lizhen,Jia Xiangxu Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing.Cloning and expression of human ciliary neurotrophic factor cDNA from fetus[J].Bulletin of the Academy of Military Medical Sciences,1998,22(2):118-121,124. |
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| Authors: | Jiang Guochun Yuan Lizhen Jia Xiangxu Institute of Radiation Medicine Academy of Military Medical Sciences Beijing |
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| Institution: | Jiang Guochun,Yuan Lizhen,Jia Xiangxu Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850 |
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| Abstract: | Objective: To clone the cDNA of human ciliary neurotrophic factor (hCNTF) and to express it in E.coli. Methods: Total RNA was extracted from cells and tissues and, after RT PCR and sequence analysis, expressed in prokaryotes and purified . Results: cDNA of CNTF was cloned and highly expressed in E.coli, with an expression level of 26%. After purification with preparative electrophoresis, the purity of the recombinant protein was 95%. Conclusion: The cDNA of CNTF was cloned from fetal sciatic nerve tissues and highly purified recombinant protein was obtained after purification with preparative electrophoresis. |
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| Keywords: | ciliary neurotrophic factor polymerase chain reaction gene expression fetus |
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