非同位素银染单链构象多态性技术筛查葡萄糖激酶基因突变  

Non isotope SSCP technique in screening glucokinase gene mutation

作　　者：李小英[1,2] 郑以漫 孙荣 朱宏达 高梅 许曼音 陈家伦 

机构地区：[1]第三军医大学附属新桥医院 [2]上海市内分泌研究所

出　　处：《中华医学检验杂志》1997年第3期163-166,共4页

摘　　要：目的建立银染单链构象多态性（ＳＳＣＰ）技术筛查葡萄糖激酶基因点突变。方法收集１０个ＮＩＤＤＭ家系的１１２例基因组（ＤＮＡ），经多聚合酶链反应（ＰＣＲ）扩增葡萄糖激酶基因第２～１０号外显子，ＳＳＣＰ电泳，硝酸银染色筛查点突变。结果发现３个家系，９例受试者第５，６号外显子出现异常ＤＮＡ片段条带，经顺序分析证实为第５号内含子１２位碱基Ｃ→Ｇ突变。结论非同位素ＳＳＣＰ技术是一种简便、经济、快速、准确和有效的筛查基因点突变的方法。Objective To establish non isotope single strand conformation polymorphism (SSCP) technique of silver staining to screen point mutation of glucokinase gene. Methods 112 samples of genome DNA from 10 NIDDM pedigrees were collected. Exon 2 10 of glucokinase gene were amplified by PCR and screened with SSCP electrophoresis and silver staining. Results Abnormal fragment bands of exon 5 and 6 were found in 9 cases of 3 pedigrees. Sequence analysis of those abnormal DNA fragments demonstrated the twelfth base replacement of C→G in intron 5. Conclusions It is concluded that SSCP technique is a simple, rapid, economic, accurate and effective method of screening gene point mutation.

关 键 词：多态现象 单链构象 葡萄糖激酶 基因突变 糖尿病 

分 类 号：R587.104[医药卫生—内分泌]