应用内含子法构建PC基因mRNA竞争RT-PCR内参照  

Construction of Competitive Reverse Transcription Polymerase Chain Reaction Inter-Reference of PC mRNA by Intron Approach

作　　者：徐闯[1] 夏成[1] 刘国文[1] 王哲[1] 姜玉富[1] 

机构地区：[1]中国人民解放军军需大学,长春130062

出　　处：《中国农业科学》2004年第7期1056-1059,共4页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助项目(30230620)

摘　　要：运用PC基因mRNA与PC基因DNA相比缺少1个81bp内含子序列的基本原理,应用PCR技术和生物工程原理,通过1对引物,进行1次克隆,成功构建了PC基因mRNA的466bp竞争DNA模板重组质粒,PC基因mRNA与PC基因DNA可以互为内参照。为应用逆转录-聚合酶链反应(RT-PCR)检测糖代谢过程中PC基因mRNA水平奠定方法学基础。Inter-reference of competitive reverse transcription polymerase chain reaction(RT-PCR) was constructed byintron method to detect the change of PC mRNA level in the pathway of carbohydrate metabolism. The experiment wasbased on the principle that 81bp intron sequence was deleted in PC mRNA compared with PC DNA sequence. The 466bpcompetitive DNA template recombinant plasmid of PC mRNA was successfully built with a pair of primer and once clone.PC DNA could be inter-reference each other. The intron approach used in experiment has broken through the traditionalmethod in building competitive template.

关 键 词：内含子法 PC基因 MRNA 竞争模板 RT-PCR 内参照 丙酮酸羟化酶 

分 类 号：Q789[生物学—分子生物学]