Screening of Reference Genes in Paenibacillus thiaminolyticus SY20 by Real-Time Fluorescene Quantitaticve PCR
In order to study the expression levels of polymyxin synthesis genes in Paenibacillus thiaminolyticus SY20,the appropriate reference gene is required to obtain reliable results in real-time quantitative polymerase chain reaction(PCR).The expressions of five candidate reference genes(RecA,RpoB,ftsZ,GAPDH and 16S rRNA)under different fermentation phases of Paenibacillus thiaminolyticus SY20 were analyzed by real-time fluorescence quantitative PCR,and their expression stabilities were evaluated by a comprehensive analysis of five software,including GeNorm,NormFinder,BestKeeper,the comparative Delta-Ct and RefFinder.The results showed that the genes of ftsZ and RecA were more stable using GeNorm analysis.But NormFinder,BestKeeper,comparative Delta-Ct and RefFinder analysis showed that ftsZ gene was the most stable.Therefore,the ftsZ gene was determined to be the most suitable reference gene for studying the gene expression of P.thiaminolyticus SY20 gene by combining five analytical methods.
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