【摘要】 目的利用限制性酶切片段长度多态性(RFLP)及变性高效液相色谱法(DHPLC)筛选和鉴定脑胶质瘤易感基因———切除修复鼠缺陷交叉互补基因2 (ERCC2)的单核苷酸多态性(SNPs)。方法应用PCR扩增179例胶质瘤病人肿瘤及血液标本和44例正常对照组血液标本ERCC2基因第23外显子及其邻近的部分内含子序列,采用限制性酶切及DHPLC技术对扩增片段进行基因变异检测,直接测序不同类型的PCR片段,并与参考序列进行对比分析。结果在此片段中验证了一个已知白种人存在的SNP位点。结论黄种人亦存在rs13181,并影响蛋白编码;可能与胶质瘤的发病有关联。DHPLC相对于RFLP来说是一种高效、经济、简便、可靠的SNPs筛选方法。更多还原

【Abstract】 Objective To screen single nucleotide polymorphisms (SNPs) of ERCC2 gene by using restriction fragment length polymorphism (RFLP) and denaturing high-performance liquid chromatography (dHPLC). Methods The sequence of 23 exons and the adjacent part of the intron in the ERCC2 gene isolating from the glioma, the blood samples of 179 glioma patients and 44 healthy controls were amplified by PCR, and the products were analyzed by RFLP and dHPLC for the gene mutation detection. Some meaningful fragments were sequenced and compared with the sequences available from National Center for Biotechnology Information (NCBI) database. Results In this study, a SNP locus which is known to exist in White patients with glioma was validated. Conclusion Gene rs13181 also exists in yellow race with the effect on protein coding, and maybe related to the occurrence of glioma. Comparing with RFLP, dHPLC is a highly effective, economical, simple and reliable method for SNPs screening.更多还原