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目的筛选和鉴定脑胶质瘤易感基因ERCC2———纠正鼠突变细胞切除修复缺陷基因的单核苷酸多态性(SNPs)。方法应用参考文献所报道的引物序列,PCR扩增179例胶质瘤患者肿瘤及血液标本和44例正常对照组血液标本ERCC2基因第22外显子及其邻近的部分内含子序列,采用DHPLC技术对扩增片段进行基因变异检测,将不同类型的PCR片段进行全序列测序并与参考序列对比分析。结果在此片段中验证了一个已知的SNP位点,初步鉴定了1个新的SNP位点,排除了1个已知在白种人存在的SNP位点和基因型。结论SNPs位点存在人种间差异;DHPLC预测结合全序列测序是一种高效、经济、简便、可靠的SNP筛选方法。
Objective To screen and identify human glioma susceptibility gene ERCC2 - to correct single nucleotide polymorphisms (SNPs) of excision and repair defects in murine mutant cells. Methods The exon 22 of ERCC2 gene and its adjacent intron sequences were amplified by PCR from 179 cases of glioma and 44 normal controls using PCR primers. DHPLC The technique was used to detect the gene mutation of the amplified fragment. The different types of PCR fragments were sequenced and compared with the reference sequence. Results A known SNP site was validated in this fragment, a new SNP site was initially identified, and one SNP site and genotype known to be present in Caucasians was excluded. Conclusion There are inter-ethnic differences in SNPs. DHPLC prediction combined with full sequence sequencing is an efficient, economical, simple and reliable SNP screening method.