期刊论文

Construction of EGFR gene G719S and T790M mutation vector and preliminary clinical application

中文

临床检验杂志

1001-764X

2018

36

5

380-383

10.13602/j.cnki.jcls.2018.05.16

湖南省自然科学基金

本校为第一机构

目的构建与宫颈癌组织表皮生长因子受体(epidermal growth factor receptor,EGFR)基因G719S和T790M位点突变型重组载体,利用其建立分子开关平台检测宫颈癌EGFR基因突变。方法以野生型重组质粒为模板,利用重叠PCR技术,得到突变型融合目的DNA片段,再将此目的片段连接入pMD19-T质粒中,构建成突变型重组载体,将其转化入大肠埃希菌E.coli DH5α感受态细胞进行表达,用菌液PCR和基因组测序进行鉴定。设计特异性检测引物,建立分子开关检测平台用于临床宫颈癌样本的检测。结果通过基因组测序证实G719S和T790M突变位点成功引入,定点突变载体构建成功。成功建立了分子开关检测平台用于宫颈癌组织DNA的检测。...

Objective To construct mutant recombinant vector of epidermal growth factor receptor ( EGFR) gene G719S and T790M sites associated with cervical cancer, lay the foundation for the detection of EGFR gene mutation in cervical cancer. And using it to es-tablish a molecular switch platform to detect cervical cancer EGFR gene mutations. Methods Using the wild-type recombinant plasmid as template, the mutant fusion target fragment were amplified by overlap PCR, then connect this target fragment into the vector pMD19-T. The constructed mutant recombin...