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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。
Objective To screen and identify the susceptibility gene of glioma by restriction fragment length polymorphism (RFLP) and denaturing high performance liquid chromatography (DHPLC) - to excise and repair a single defect of murine defect cross-complementing gene 2 (ERCC2) Nucleotide polymorphisms (SNPs). Methods The exon 23 of ERCC2 gene and its adjacent intron sequences were amplified by PCR from 179 glioma patients and 44 normal controls. The restriction endonuclease digestion and DHPLC An amplified fragment was used to detect the gene mutation. Different types of PCR fragments were directly sequenced and compared with the reference sequence. Results In this fragment, a known Caucasian SNP site was validated. Conclusion There are also yellow rs13181, and affect the protein coding; may be associated with the pathogenesis of glioma. DHPLC relative to RFLP is an efficient, economical, simple and reliable SNPs screening method.