实时荧光定量PCR分析家蚕杆状病毒中多角体蛋白基因(polh) mRNA的半衰期
Real-time quantified PCR analysis of half-life of polynedron gene(polh) mRNA in Bombyx mori nuclear polyhedrosis virus
家蚕多角体蛋白是一种杆状病毒高效表达的蛋白质，为了研究其mRNA的半衰期，我们首先用放线菌素D阻断polh基因的转录，然后在不同时间点分别提取RNA后再利用实时荧光定量PCR检测polh mRNA的含量和半衰期；通过实验我们得出polh的mRNA半衰期为12h，放线菌素D阻断后2 h和4 h时mRNA降解程度不大（p>0.05），8 h和12 h时降解显著（p<0.05）和极为显著（p<0.01）。多角体蛋白基因polh的mRNA半衰期比目前已知的其他基因mRNA半衰期较长，这可能和蛋白的高效表达存在着一定的关系，也为进一步研究多角体蛋白高效表达提供了一种新的研究途径。
Bombyx mori polyhedrin(Polh) is a high expressed protein in baculovirus. In order to study the half-life of polh mRNA connected with high expression , we firstly used actinomycin D(act D) to blocking-up polh gene transcription, and secondly extracted total RNA of virus and cells at different times. Thirdly, we detected mRNA copies and half-life of polh by real-time quantified PCR. The result is that the half-life of polh mRNA is 12 h , and the mRNA degradate a little at 2 h and 4 h (p>0.05)and significantly at 8 h (p<0.05)and 12 h (p<0.01). The half-life of polh is longer than other gene mRNA that had been known, and it may have a connection with high expression. It also provide a new way to the high expression study of Polh..
耿文杰、张耀洲、舒特俊、解纯刚、陈剑清
分子生物学昆虫学
分子生物学实时荧光定量PCR多角体基因(polh)mRNA半衰期多角体高效表达
Molecular BiologyReal time quantified PCRPolynedron gene(polh)mRNAHalf-lifePolynedron high expression
耿文杰,张耀洲,舒特俊,解纯刚,陈剑清.实时荧光定量PCR分析家蚕杆状病毒中多角体蛋白基因(polh) mRNA的半衰期[EB/OL].(2012-12-27)[2025-07-21].http://www.paper.edu.cn/releasepaper/content/201212-999.点此复制
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