改进竞争性RT-PCR法在定量PER1基因表达中的应用  

Improvement of competitive RT-PCR in quantification of PER1 gene expression

作　　者：刘姝[1] 蔡彦宁[1] 冯秀丽[1] 温玫[1] 左晓虹[1] 陈彪[1] 

机构地区：[1]首都医科大学宣武医院老年病医学研究所神经生物学室,北京100053

出　　处：《基础医学与临床》2005年第5期409-413,共5页Basic and Clinical Medicine

基　　金：国家科技攻关计划(2002BA711A07-04);国家重点基础研究计划(973)(G2000057005);北京市自然科学基金重点项目(7031002);北京市科技新星项目(H020821400190);北京市卫生局重点学科项目

摘　　要：目的研究使用不同样品稀释液及变性高效液相色谱(DHPLC)梯度条件对定量检测PER1基因表达的影响。方法竞争性RT-PCR与DHPLC结合定量检测基因表达。结果样品稀释液中含有载体可极大提高实验的重复性、准确性;不同DHPLC梯度条件对定量结果无明显影响。结论建立定量PER1基因mRNA表达水平的竞争性RT-PCR系统有助于精确检测微量标本中节律基因表达的变化,探寻生物节律与疾病状态间的关系。ObjectiveTo study the impact of different dilution solution and DHPLC washing condition on quantification. MethodsCompetitive RT-PCR system based on DHPLC was used in order to detect PER1 gene expression. ResultsAddition of carriers significantly improved the accuracy and reproducibility of the experiment. Different washing conditions of DHPLC had no obvious influence on the quantification. ConclusionEstablishment of this system would contribute to the study of circadian rhythm in tissues and explore the relationship between circadian rhythm and diseases.

关 键 词：PCR法 竞争性RT-PCR 变性高效液相色谱 基因MRNA表达 DHPLC 检测基因表达 定量检测 CR系统 疾病状态 生物节律 稀释液 重复性 准确性 梯度 样品 

分 类 号：R346[医药卫生—基础医学]