高效毛细管电泳与聚丙烯酰胺凝胶电泳检测基因点突变的比较
The comparison of detection of gene point mutation by HPCE-SCP and PAGE-SCP analysis
作者：刘戟;王若菡;茹钦华;
Author：
收稿日期： 年卷（期）页码：2006,(04):-342-344
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：毛细管电泳;聚丙烯酰胺凝胶电泳;p53基因;点突变
Key words：
基金项目：
中文摘要
目的比较高效毛细管电泳(HPCE)技术和传统聚丙烯酰胺凝胶电泳(PAGE)检测结肠癌DNA是否存在p53基因点突变的方法。方法分别采用HPCE与PAGE作单链构象多态性(SCP)分析,检测结肠癌肿瘤标本p53基因第7外显子PCR扩增产物,DNA测序检测其准确性。结果利用HPCE技术和PAGE结合银染技术从22例结肠癌标本中分别检测出5例和4例突变,测序证实点突变的确存在。结论HPCE技术较PAGE更为快速、简便和准确,对临床筛查基因点突变有较高的应用价值。
参考文献
[1]林炳承.毛细管分析导论[M].北京:科学出版社,1996,1-346.
[2]P ier G iorgio R ighetti,Cecilia Gelfi.Cap illary electrophoresis ofDNA for molecu lar d iagnostics[J].E lectrophoresis,1997,18:1709-1714.
[3]Keith R.M itchelson,Jing Cheng,larry J.Kricka.The Use of cap-illary electrophoresis for point-mutation screen ing[J].Tibtech,1997,15:448-458.
[4]Felm lee TA,Oda RP,Persing DA,et a.l Cap illary electrophoresisof DNA:Potential utility for clin ical d iagnoses[J].J ChromatA,1995,717:127-137.
[5]Sambrook J,Fritsch EF,Man iatis T.Molecu lar clon ing:A labora-tory manual[M].New York:Cold Spring Harbor LaboratoryPress,1989,174.
[6]Dockorn Dworn iczak B.Non istop ic detection of single-strandconformation polymorph ism:A rap id techn ique in d iagnosis ofphenyketonnuria[J].Nucleic Acids Res,1991,19:2500-2502.
[7]A ldy WH Kuypers,Peter MW W illem s,Ewald JBM M ensink,etal.D etection of point mutations in DNA using cap illary electro-phoresis in a polym er network[J].J Chromat,1993,621:149-156.
[8]H idetosh i Arakawa,Shu ji Nakash iro,MasakoMaeda,et al.Analy-sis of single-strand polymorph ism by cap illary electrophoresis[J].J ChromatA,1996,722:359-368.
[9]Donald H Atha,M ichaelW enz H,Harvard Morehead,et al.D e-tection of p53 pointmutation by single strand conformation poly-morph ism:Analysis by cap illary electrophoresis[J].E lectropho-resis,1998,19:172-179.
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