毛细管电泳方法快速检测肺癌组织p53基因第249 位密码子点突变研究
首发时间:2008-04-10
摘要:目的:p53基因点突变在肺癌的发生过程中起重要作用,检测基因点突变的方法学研究将有助于临床准确诊断肺癌。方法:运用毛细管电泳(CE)与聚丙烯酰胺电泳(PAGE)两种方法对肺癌及癌旁正常组织p53基因第249位密码子点突变进行检测,并对其检测结果进行比较。PAGE以15%聚丙烯酰胺作为分离胶;毛细管电泳分离条件为2.0%甲基纤维素(MC)(pH=8.0)筛分介质,分离温度25℃,电场强度200V/cm。结果:用毛细管电泳和聚丙烯酰胺凝胶电泳两种方法均能检测到p53基因第249位密码子点突变,而且毛细管电泳方法检测时间短,且检测率高。结论:毛细管电泳可为大规模进行肺癌的早期诊断提供简便可靠的方法。
关键词: 毛细管电泳 限制性片段长度多态性 p53基因 点突变
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Fast Detection of the 249 coden of p53 Gene point Mutation of lung Cancer by Restriction Fragment Length Polymorphism With Capillary Electrophoresis
Abstract:Objective:p53 gene point mutation is important in lung cancer. Compare the result of the 249 coden of p53 Gene point Mutation by Polyacrylamide Gel Electrophoresis(PAGE) and Capillary Electrophoresis(CE).Methods:Detect the 249 coden of lung cancer and normal lung tissue p53 Gene point Mutation by PAGE and CE simultaneously. The separation gel of PAGE is 15% Polyacrylamide Gel. The condition of CE was 2.0%(pH=8.0) Methyl Cellulose(MC),separation temperature at 25℃,electric field strength at 200V/cm.Results:The CE and PAGE can both detect the 249 coden of p53 Gene point Mutation. The detection time of CE is short,and detection rate is high.Conclusion:CE can offer a convenient and reliable method to the large scale of early diagnosis of lung cancer.
Keywords: Capillary Electrophoresis;Restriction Fragment Length Polymorphism;p53 gene;point mutation
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毛细管电泳方法快速检测肺癌组织p53基因第249 位密码子点突变研究
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