Screening of Reference Genes in Bacillus pumilus by Real-time Fluorescence Quantitative PCR

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.028

Abstract

Abstract: In order to study the gene expression in Bacillus pumilus through real-time fluorescence quantitative PCR,we need to determine the appropriate reference gene firstly. The expressions of five candidate reference genes（16S rRNA,mecA,cadR,rpoB,andsphP）under different fermentation phases of B. pumilus were analyzed by real-time fluorescence quantitative PCR,and their expression stabilities were evaluated by geNorm and NormFinder software. According to the analysis by geNorm software,16S rRNA and mecA were the most stable genes and the number of optimal reference genes were 2. Analysis by NormFinder software revealed that mecA was the most stable gene. The differential expressions of 3 functional genes of B. pumilus indicated that 16S rRNA and mecA were appropriate reference genes,moreover,expression abundance of mecA was moderate,thus it was more suitable to be used as a reference gene in studying structural gene. Additionally,2 reference genes can also be used for calibration in order to acquire more accurate results.

Key words: Bacillus pumilus, real-time fluorescence quantitative PCR, reference gene, geNorm, NormFinder

HE Ting-ting, SONG Ting, WANG Chao, ZHANG Chang-bin, WANG Hai-yan. Screening of Reference Genes in Bacillus pumilus by Real-time Fluorescence Quantitative PCR[J]. Biotechnology Bulletin, 2016, 32(11): 99-106.

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