一种快速获得基因密码子偏爱性改造的方法——SOE-PCR  

A Rapid Method for Generation of Codon-modifications—SOE-PCR

作　　者：王艳君[1] 杨谦[1] 

机构地区：[1]哈尔滨工业大学生命科学与工程系,哈尔滨150001

出　　处：《中国生物工程杂志》2008年第8期96-99,共4页China Biotechnology

基　　金：国家"863"计划(2003AA241140);黑龙江省国际科技合作项目(WC05B04);黑龙江省自然科学基金重点项目(ZJN03-04)资助项目

摘　　要：SOE-PCR采用具有互补末端的引物,使PCR产物形成重叠链,在不需要内切酶消化和连接酶处理的条件下实现DNA片段的拼接,能够高效、快速地实现基因的定点突变。应用SOE-PCR原理成功地实现了对几丁质酶基因chi58的5个位点的突变,构建了几丁质酶基因密码子偏爱性改造突变体—chi58A。实验证明,SOE-PCR具有简捷、快速、高效的优点。扩增过程中未引入其他突变,获得的chi58A突变体基因片段可以用于后续的分子克隆。SOE PCR, which employs chimeric primers to generate PCR products with complementary ends in amplifications. This can assemble DNA fragments without the treatment of restriction endonucleases and T4 DNA ligase. Five site-directed mutagenesis of chitinase （chi58） were obtained successfully with SOE-PCR. The mutation of codon-modifications of chi58 was constructed. The results showed that SOE-PCR have the characters of simplification and high-efficiency. Other mutations were not caused during the progress of amplification. The mutation of chi58A can be used at following research.

关 键 词：SOE—PCR 多点突变 密码子偏爱性改造 

分 类 号：Q254[生物学—细胞生物学]