| 斑马鱼TK1基因的克隆表达 |
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| 引用本文： | 杜长青,初金鑫,赵春玲,刘顺梅.斑马鱼TK1基因的克隆表达[J].生物技术,2012,22(2):1-4. |
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| 作者姓名： | 杜长青 初金鑫 赵春玲 刘顺梅 |
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| 作者单位： | 潍坊医学院药学与生物科学学院,山东潍坊,261053 |
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| 摘 要： | 目的:克隆斑马鱼TK1基因的cDNA序列,并在大肠杆菌中诱导表达,对其产物进行生物学活性鉴定。方法:采用RT-PCR和RACE方法,克隆TK1的cDNA全长序列。表达载体在大肠杆菌BL21(DE3)中进行诱导表达。表达蛋白利用镍离子柱纯化。结果:获得TK1基因的cDNA全长序列,编码一个分子量为26kD的蛋白。结论:TK1融合蛋白在28℃条件表现出比较高的生物学活性,达到0.45 U/mg。
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| 关 键 词： | 斑马鱼 胸苷激酶 克隆表达 生物活性 |
Cloning and Expression of Thymidine Kinase from Zebrafish Danio rerio |
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| DU Chang-qing
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CHU Jin-xin
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ZHAO Chun-ling
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LIU Shun-mei.Cloning and Expression of Thymidine Kinase from Zebrafish Danio rerio[J].Biotechnology,2012,22(2):1-4. |
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| Authors: | DU Chang-qing CHU Jin-xin ZHAO Chun-ling LIU Shun-mei |
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| Institution: | (Pharmacy and Biological Science College,Weifang Medical University,Weifang 261053,China) |
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| Abstract: | Objective: To clone the TK1 cDNA sequence of zabrafish,to express its protein in E.coli and to evaluate its biological activity.Method: The complete TK1 cDNA was amplified by RT-PCR and RACE.Expression vectors were induced by IPTG in E.coli BL21(DE3).TK1 protein was purified to homogeneity using Ni-NTA spin column.Result: The complete TK1 cDNA sequence was cloned,encoding a protein with a molecular weight of 26 kD.Conclusion: The purified zebrafish TK1 showed the highest biological activity,0.45 U/mg at 28℃. |
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| Keywords: | zebrafish thymidine kinase cloning and expression biological activity |
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