Selection of Reference Genes in Saccharopolyspora Spinosa for Real-Time PCR  

作　　者：张传波 薛超友 申月琪 卢文玉 

机构地区：[1]School of Chemical Engineering and Technology, Tianjin University [2]Key Laboratory of System Bioengineering(Tianjin University), Ministry of Education [3]SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering(Tianjin)

出　　处：《Transactions of Tianjin University》2015年第5期461-467,共7页天津大学学报（英文版）

基　　金：Supported by the National Natural Science Foundation of China(No.21076148 and 31270087);Plan for Tianjin Science and Technology Support(No.11ZCKFSY0100)

摘　　要：Reverse transcription quantitative PCR （RT-qPCR） combined with the published genome information of Saccharopolyspora spinosa can allow sophisticated studies about S. spinosa, including Studying the regulation of spinosyn biosynthesis, finding new target genes for engineering, and discovering and exploiting other macrolide secondary metabolites. Studies have demonstrated that appropriate internal control is needed to normalize target genes at transcription levels. However, many studies have shown that no single reference gene is universal for all strains under all experimental conditions. Thus, eight candidate reference genes of three different S. spinosa strains in two different cultures were studied to find suitable reference gene（sl. The number of amplification cycles of these candidate genes was calculated by BestKeeper, NormFinder and geNorm. The results indicated that the most suitable reference genes for normalization during the fermentation of S. spinosa were 16S rRNA and rbL13.Reverse transcription quantitative PCR(RT-q PCR)combined with the published genome information of Saccharopolyspora spinosa can allow sophisticated studies about S. spinosa, including studying the regulation of spinosyn biosynthesis, finding new target genes for engineering, and discovering and exploiting other macrolide secondary metabolites. Studies have demonstrated that appropriate internal control is needed to normalize target genes at transcription levels. However, many studies have shown that no single reference gene is universal for all strains under all experimental conditions. Thus, eight candidate reference genes of three different S. spinosa strains in two different cultures were studied to find suitable reference gene(s). The number of amplification cycles of these candidate genes was calculated by Best Keeper, Norm Finder and ge Norm. The results indicated that the most suitable reference genes for normalization during the fermentation of S. spinosa were 16 S r RNA and rb L13.

关 键 词：real-time PCR reference genes SaecharopOlysporaspinosa 

分 类 号：Q933[生物学—微生物学]