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  

Constructing Human Mutant IDUA Gene by in Vitro Site-Directed Mutagenesis Technique

作　　者：于洪枫[1] 曾瑞萍[1] 葛春喜[1] 林群娣[1] 

机构地区：[1]中山医科大学医学遗传教研室,广东广州510089

出　　处：《中山医科大学学报》2002年第2期105-107,共3页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：国家教委博士点基金资助项目 (NO2 0 0 0 0 44)

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。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.

关 键 词：定点诱变 α-L-艾杜糖醛酸酶 基因 IDUA 体外表达 基因突变 

分 类 号：Q75[生物学—分子生物学]