期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

重亚硫酸盐修饰直接测序技术检测p16基因甲基化的研究 被引量：2

Methylation detection of p16 gene using directed PCR product sequencing
下载PDF
导出
摘要 目的建立稳定的重亚硫酸盐直接测序技术,检测p16基因甲基化状况。方法提取正常人全血基因组DNA,建立起稳定的重亚硫酸盐直接测序平台,利用该技术检测结直肠癌细胞株p16基因甲基化状况。结果应用列联表的Fisher,ExactTest比较3种纯化方法的测序结果,乙醇/醋酸钠和过柱纯化与SAP-ExonI纯化法比较,测序结果差异有统计学意义(P<0.05);应用重亚硫酸盐直接测序技术,可检测出p16基因目的片段中CpG岛所有CpG位点的甲基化状况。结论应用SAP-ExonI纯化法,建立了稳定的重亚硫酸盐直接测序技术,并可检测出目的片段中CpG岛所有CpG位点的甲基化状况。 Objective To construct method of bisulfite-modifieation directed PCR product sequencing,and to identify methylation of p16 gene. Methods Ten PCR samples were purified by spin-column,ethanol/ sodium acetate and SAP-Exonl. Results The sequencing results of SAP Exonl purification method were better than those of the other two methods significantly（P〈50.05）. Conclusion Satisfactory results can be got when use SAP-ExonI purification method.
作者 张华 府伟灵 黄庆 郭颖
机构地区 第三军医大学西南医院检验科
出处 《重庆医学》 CAS CSCD 2006年第3期222-223,共2页 Chongqing medicine
基金 贵州省科技攻关资助项目(黔科合NY字[2005]3003号)
关键词 双脱氧测序法 虾碱性磷酸酶(SAP) 核酸外切酶I(EXONI) 测序图谱 sanger dideoxy DNA sequencing sequencing chromatogram EXON I （ exonuclease I） SAP （ shrimp alkaline phosphatase）
分类号 R735.3 [医药卫生—肿瘤]
  • 相关文献

参考文献6

  • 1Lin SY, Yeh KT, Chen WT, et al. Promoter CpG methylation of tumor suppressor genes in colorectal cancer and its relationship to clinical features[J]. Oncol Rep,2004,11(2):341.
  • 2Tycko B. Epigenetic gene silencing in cancer[J]. J Clin Invest,2000,105(4) :401.
  • 3孙念绪,林武华,蔡志民.错配修复、微卫星不稳定性与散发性结直肠癌[J].重庆医学,2002,31(4):269-270. 被引量：3
  • 4赵晓晏,尚瑞莲,夏时海,郭萍.原发性胰腺癌中p16基因突变及其蛋白表达的研究[J].重庆医学,2002,31(3):163-166. 被引量：8
  • 5Stirzaker C, Song JZ, Davidson B, et al. Transcriplional gene silencing promotes DNA hypermethylation through a sequential change in chromatin modifications in cancer cells[J]. Cancer Res, 2004,64 : 3871.
  • 6Frommer M , McDonald DS, Millar CM ,et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands [J].Proc Natl Acad Sci,1992,89:1827.

二级参考文献12

  • 1Leach FS,Nicholas C,Fredrick S,et al.Mutation of a muts homology in hereditary nonpolyposis colorectal cancer[].Cell.1993
  • 2Reenan RAG,Kolodner RD.Isolation and characterization of two saccharo wycas cerevisiae genes encoding homologous of the bacterial HexA and muts mismatch repairprotein[].Genomics.1992
  • 3Lindor NMJ,Jalal SM,Van Dawlker,et al.Search of chromosome instability in lymphocytes with germ-line mutation in DNA mismatch repair genes[].Cancer Genetics and Cytogenetics.1998
  • 4Aaltonen LA,Thibodeau G,Bren D,et al.Microsatellite instability in cancer of the proximal colon[].Science.1993
  • 5Frideberg EC.The enzymology of DNA repair[].Mutation Research.1990
  • 6Fishel,Lescoe MK,Rao MRS,et al.The human mutation gene homology MSH2 and its association with hereditary nonpolyposis colon cancer[].Cell.1993
  • 7Wagner R,Meseison H.Repair tractsin mismatched DNA heteroduplexes[].Proceedings of the National Academy of Sciences of the United States of America.1996
  • 8Aaltonen LA,Lauri A,Leach,et al.Clues to the pathogenesis of familial colorectal cancer[].Science.1993
  • 9Keller G,Rudelious M,Vogelsang H,et al.Genomice instability in colorectal cancer: relationship to clinicopathological variables and family history[].Cancer Research.1998
  • 10Aaltonen LA,Zauri A,Jukka-pekka,et al.Replication error in benign and maligant tumors from hereditary nonpolyposis colorectal cancer patients[].Cancer Research.1994

共引文献9

同被引文献26

  • 1Clark S J, Harrison J, Paul C L, et al. High sensitivity mapping of methylated cytosines[ J]. Nucleic Acids Res, 1994, 22(15) :2990-2997.
  • 2Monk M. Changes in DNA methylation during mouse embryonic development in relation to X-chromosome activity and imprinting [ J ]. Philos Trans R Soc Lond B Biol Sci, 1990,326(1235) :299-312.
  • 3Howlett S K, Reik W. Methylation levels of maternal and paternal genomes during preimplantation development [ J ]. Development, 1991,113 ( 1 ) : 119-127.
  • 4Frommer M, McDonald L E, Millar D S,et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands [ J ]. Proc Natl Acad Sci USA, 1992,89 ( 5 ) : 1827-1831.
  • 5Olek A, Oswald J, Walter J. A modified and improved method for bisulphate based cytosine methylation analysis [ J]. Nucleic Acids Res, 1996,24(24) :5064-5066.
  • 6Hajkova P, el-Maarri O, Engemann S, et al. DNA-methylation analysis by the bisulfite-assisted genomic sequencing method [ J ]. Methods Mol Biol, 2002,200 : 143-154.
  • 7Yamazaki Y, Fujita T C, Low E W, et al. Gradual DNA demethylation of the Oct4 promoter in cloned mouse embryos [ J ]. Mol Reprod Dev, 2006,73 (2) : 180-188.
  • 8Chen T, Zhang Y L, Jiang Y, et al. Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos [ J ]. Mol Reprod Dev,2006,73 (3) :313-317.
  • 9Gomyo H,Arai Y,Tanigami A,et al.A 2-Mb sequenceready contig map and a novel immunoglobulin superfamily gene IGSF4 in the LOH region of chromosome 11q23.2[J].Genomics,1999,62(2):139.
  • 10Kuramochi M,Fukuhara H,Nobukuni T,et al.TSLC1 is a tumor-suppressor gene in human non-small-cell lung cancer[J].Nature Genetics,2001,27(4):427.

引证文献2

二级引证文献2

重庆医学
2006年 第3期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部