非同位素PCR－单链构象多态性技术的建立和应用

ＰＣＲ－单链构象多态性技术问世以来，成为研究基因突变的工具，特别是在分子肿瘤学研究中，广泛应用于癌基因，抑癌基因突变的研究，常规ＰＣＲ－ＳＳＣＰ采用同位素标记ＰＣＲ产物，测序板电泳分离突变，在操作和费用上有种种局限，文章建立了一种非同位素ＰＣＲ－ＳＳＣＰ技术；通过不对称ＰＣＲ获得单链，普通ＰＡＧＥ分离，经银染检出突变，用这种方法，还研究了四株鼻咽癌细胞株ＣＮＥ１，ＣＮＥ２，ＨＫ１和ＳＵＮＥ１中肿瘤

A Non-radioactive Single-strand Conformation Polymorphisms of Asymmetric PCR ans its Application

PCR-single strand conformation polymorphisms is a powerful method for screening mutations and widely used in studying mutations of oncogene and tumor suppressor gene.The ordinary PCR-SSCP needs the use of radioactivity and sequencing appratus, thus compromises its application.Here,a non-radioaltive asymmetric PCR-SSCP was established.Single-stranded DNA was generated by asymmetric PCR, seperated by mini PAGE and silver stained. The exon 5,6,7,8 of p53 gene in four cell lines of nasopharyngeal carcinoma-CNE1, CNE2, HK1 and SUNE1 were investigated. The method was proved to be succssful in screening mutations.