摘要
目的:建立一种半定量RT-PCR方法,用于齿肋赤藓(Syntrichia caninervis)基因表达研究。方法:比较常用的几种内参照基因actin、tubulin及18S rRNA在齿肋赤藓中的表达情况,以确定表达稳定的内参基因;分析PCR扩增动力学,确定内参和靶基因的PCR体系。结果:琼脂糖凝胶电泳检测分析表明:18S rRNA作为齿肋赤藓的内参基因更稳定;同管扩增试验表明:18SrRNA的引物滞后6个循环加入PCR扩增体系中,至第26个循环,二者同时处于指数扩增期,且靶基因RT-A的扩增效率不受影响。结论:18S rRNA与RT-A可以同管扩增,18S rRNA可以作为半定量RT-PCR的内参照,为齿肋赤藓基因表达研究奠定基础。
Objective：Make sure a kind of method of Semi-Quantitative RT-PCR,so that it can be used to analyze the gene expression in Syntrichia caninervis.Method：Compared the expression of some usual internal standard genes,containing actin,tubulin and 18S rRNA and make sure the internal standard gene in Syntrichia caninervis.At the same time,analyzed the PCR dynamics and make sure the PCR system.Result：The analysis of agarose gel electrophoretic shows 18S rRNA as internal standard gene is more constant.The experiment of amplification in the same tube reveals that the primes of 18S rRNA as internal standard gene were added after 6 cycles that of interesting gene Internal standard gene,when the 26th cycles,they arrived the exponential period at the same time,and the PCR efficiency of RT-A is not decline.Conclusion：18S rRNA can be amplified with targeted gene in the same tube.The 18S rRNA can be as internal standard,the research provide theoretic reference in studying gene expression in Syntrichia caninervis.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第2期23-25,共3页
Biotechnology
基金
新疆自治区重大专项(200731138-3)
国家重点基础研究发展计划(2009CB825104)
农业部转基因科技重大专项(2009zx08005-022B)资助
