| 利用体外定点诱变技术构建人突变型α-L-艾杜糖醛酸酶基因 |
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| 引用本文： | 于洪枫,曾瑞萍,葛春喜,林群娣.利用体外定点诱变技术构建人突变型α-L-艾杜糖醛酸酶基因[J].中山大学学报(医学科学版),2002,23(2):105-107. |
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| 作者姓名： | 于洪枫 曾瑞萍 葛春喜 林群娣 |
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| 作者单位： | 中山医科大学医学遗传教研室,广东,广州,510089 |
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| 基金项目： | 国家教委博士点基金资助项目 (NO2 0 0 0 0 44) |
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| 摘 要： | 【目的】构建突变型α L 艾杜糖醛酸酶基因 (IDUA)cDNA ,为体外表达、鉴定突变的性质提供物质基础。【方法】以野生型pcDNA3 IDUA为基础质粒 ,采用双引物法体外定点诱变技术 ,引入突变E40 4X。【结果】突变区域经PCR SSCP和测序证实诱导突变成功。【结论】本研究成功构建了突变型IDUAcDNA ,可用于下游的体外表达 ;此定点突变技术具有简便、诱变效率高等优点 ,是体外构建突变型基因的一种有效方法。
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| 关 键 词： | 诱变 定点 α-L-艾杜糖醛酸酶 基因 IDUA 突变 |
| 文章编号： | 1000-257X(2002)02-0105-03 |
| 修稿时间： | 2001年10月11 |
Constructing Human Mutant IDUA Gene by in Vitro Site-Directed Mutagenesis Technique |
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| YU Hong-feng,ZENG Rui-ping,GE Chun-xi,LIN Qun-di..Constructing Human Mutant IDUA Gene by in Vitro Site-Directed Mutagenesis Technique[J].Journal of Sun Yatsen University(Medical Sciences),2002,23(2):105-107. |
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| Authors: | YU Hong-feng ZENG Rui-ping GE Chun-xi LIN Qun-di |
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| Abstract: | Objective The mutant IDUA gene (IDUA) cDNA was constructed in order to establish the foundation for in vitro expression and further identification. Methods Using double primer in vitro site-directed mutagenesis, the mutation was induced into the wild IDUA cDNA. Results The mutant IDUA cDNA gene was successfully constructed, which was proved by PCR-SSCP and sequencing. Conclusion The mutant IDUA cDNA is going to be used for further analysis of the mutation; this site-directed mutagenesis technique has the advantages of simplicity and high-efficiency. |
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