Reliable reference gene selection for Cordyceps militaris gene expression studies under different developmental stages and media----中国科学院微生物研究所

科研成果

论文题目:	Reliable reference gene selection for Cordyceps militaris gene expression studies under different developmental stages and media
作者:	Lian Tiantian, Yang Tao, Liu Guijun, Sun Junde, Dong Caihong*
联系作者:	Dong Caihong*
刊物名称:	FEMS Microbiol Lett
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年份:	2014
影响因子:	2.286
论文下载:	http://onlinelibrary.wiley.com/doi/10.1111/1574-6968.12492/abstract
摘要:	Cordyceps militaris is considered a model organism for the study of Cordyceps species, which are highly prized in traditional Chinese medicine. Gene expression analysis has become more popular and important in studies of this fungus. Reference gene validation under different experimental conditions is crucial for RT-qPCR analysis. In the present study, eight candidate reference genes, actin, cox5, gpd, rpb1, tef1, try, tub and ubi, were selected and their expression stability was evaluated in C. militaris samples using four algorithms, GeNorm, NormFinder, BestKeeper and the comparative Ct method. Three sets of samples, five different developmental stages cultured in wheat medium and pupae, and all the samples pool were included. The results showed that rpb1 was the best reference gene during all developmental stages examined, while the most common reference genes, actin and tub, were not suitable internal controls. Cox5 also performed poorly and was less stable in our analysis. The ranks of ubi and gpd were inconsistent in different sample sets by different methods. Our results provide guidelines for reference gene selection at different developmental stages and also represent a foundation for more accurate and widespread use of RT-qPCR in C. militaris gene expression analysis. This article is protected by copyright. All rights reserved.