非同位素PCR－单链构象多态性技术的建立和应用

A Non-radioactive Single-strand ConformationPolymorphisms of Asymmetric PCR ans its AP-plication.

ＰＣＲ－单链构象多态性技术（ＳＳＣＰ）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中肿瘤抑制基因ｐ５３基因突变。证实ＣＮＥ１，ＣＮＥ２在ｅｘｏｎ８，ＨＫ１在ｅｘｏｎ５有突变，并发现新建立的细胞株ＳＵＮＥ１在ｅｘｏｎ８有突变。

CR-single strand conformation polymor-phisms is a powerful method for screening mu-tations and widely used in studying mutationsof oncogene and tumor suppressor gene.Theordinary PCR-SSCP needs the use of radioac-tivity and sequencing appratus, thus compro-mises its application.Here,a non-radioaltiveasymmetric PCR-SSCP was established.Sin-gle-stranded DNA was generated by asymmet-ric PCR, seperated by mini PAGE and silverstained. The exon 5,6,7,8 of p53 gene infour cell lines of nasopharyngeal carcinoma-CNE1,CNE2,HK1 and SUNE1 were investi-gated. The method was proved to be succssfulin screening mutations.