期刊论文

Detection of the gene mutations of lS-thalassemia by gene mutation-sensitive molecular switch

中文

实用医学杂志

1006-5725

2012

28

2

178-180

10.3969/j.issn.1006-5725.2012.2.006

国家高技术研究发展计划（编号:863计划2008AA02Z436）；

本校为其他机构

目的：采用高保真DNA聚合酶介导的基因突变敏感性分子开关准确、快速检测B地中海贫血。方法：首先提取正常人血液基因组DNA，根据已知8珠蛋白基因热点突变CD41—42、IVS2．654区域序列．分别设计突变序列扩增引物，利用低保真酶进行引物延伸反应，将其PCR产物克隆到pMDl9一T载体，得到B地中海贫血常见的两个突变位点的基因突变克隆：CD41．42、IVS2—654，然后以这两个突变位点为检测靶点，分别设计3’末端与野生型基因位点或突变基因位点配对的引物．硫化磷酸修饰3’末端并偶联高保真DNA聚合酶构成的基因突变敏感性分子开关．对含有相应突变的阳性质粒模板及正常人基因组DNA模板进行引物延伸反...

Objective To detect β-thalassemia by the approach of gene mutation-sensitive molecular switch. Methods Genomic DNA was extracted from normal blood. According to the sequences of known hot spot mutation region of CD41-42 and IVS2-654 inβ-globin gene, the mutant sequence primer was designed, and then primer extension reaction was performed with low-fidelity enzyme, and the PCR products was cloned into pMD19-T vector. Two kinds of clones of 13-thalassemia mutation gene （CD41-42 and IVS2-654） were obtained. Then these two mutation region were u...