用竞争性RT-PCR方法定量检测p53基因的表达

Quantitative determination of p53 gene expression by competitive RTPCR

目的：建立定量检测ｐ５３基因表达的方法。方法：采用竞争性逆转录聚合酶链反应（ＲＴ－ＰＣＲ）方法定量检测了组织和细胞内抑癌基因ｐ５３ｍＲＮＡ含量的变化，同时用狭线杂交的方法进行了对照。结果：竞争性ＲＴ－ＰＣＲ和狭线杂交均反映了ｐ５３基因表达变化的趋势，两者的结果相吻合。与狭线杂交相比，竞争性ＲＴ－ＰＣＲ具有灵敏度高、专一性好、简便、快速等特点，尤适合于定量检测微量样品和表达水平较低的基因。结论：竞争性ＲＴ－ＰＣＲ是一种较好的定量检测基因表达水平的方法。

Objective: To establish a method for quantitative determination of p53 gene expression. Methods: The levels of p53 mRNA in tissues and cells were investigated by competitive reverse transcription polymerase chain reaction (RTPCR) and slot blot. Results: The level of p53 gene expression measured by competitive RTPCR was similar to that by slot blot. In comparision with slot blot, competitve RTPCR was more sensitive, specific and rapid, especially suitable for microsample and the genes of low expression. Conclusion: competitive RTPCR may be a good method for quantitative determination of gene expression.