限制酶切指纹-单链构象多态性——一种有效的大片段中突变的检测方法  

Restriction endonucleases fingerprinting-single strand conformation polymorphism:an efficient method to screen mutations in long segments

作　　者：任鲁风[1] 徐悦凡[1] 杨宇[2] 

机构地区：[1]天津市妇女保健所遗传室,300204 [2]南开大学染色体实验室

出　　处：《中华医学遗传学杂志》2002年第5期416-419,共4页Chinese Journal of Medical Genetics

基　　金：天津市卫生局科技基金 (99KY0 4 4 )~~

摘　　要：目的　建立一种简便、价廉、有效地在大片段中检测突变的限制酶切指纹 -单链构象多态性(restriction endonuclease fingerprinting- single strand conformation polymorphism,REF- SSCP)方法。方法　以耳聋患者基因组 DNA为模板 ,扩增 Cx2 6基因片段 ,通过对扩增产物进行单链构象多态性、限制酶切指纹 -单链构象多态性和 DNA测序 ,比较 REF- SSCP技术对大片段中突变的检出效率。结果　长度为 72 4bp的扩增产物 SSCP结果未显示差异 ,REF- SSCP显示 3种不同带型 ,经测序发现该基因中的 79G→ A突变 ,突变情况与 REF- SSCP带型完全吻合 ,检出率为 10 0 %。结论　所建立的 REF- SSCP技术适用于大样本量地检测大片段Objective To develop a simple, cheap and efficient restriction endonucleases fingerprinting single strand conformation polymorphism(REF SSCP) method applied to screen for mutations in long segments. Methods The genomic DNA of Cx26 gene segment of the patients with deafness was amplified. The amplification products were screened with SSCP and REF SSCP technique and DNA sequencing to evaluate and compare the effect on detection of mutations in long segments. Results No different band was found in 724 bp segment in SSCP examination. Three kinds of different bands were discovered in REF SSCP examination and the 79 G→A mutation detected by DNA sequencing were accorded with the REF SSCP bands entirely. The rate of detection was 100%. Conclusion The present REF SSCP method is applicable to screen mutations in long segment DNA of mass specimens.

关 键 词：限制酶切指纹-单链构象多态性 Cx26基因 突变检测 基因突变 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]