Validation of Reference Genes for Real-Time Quantitative PCR (qPCR) Analysis of Avibacterium paragallinarum

被引：23

作者：

Wen, Shuxiang ^{[1
]}

Chen, Xiaoling ^{[1
]}

Xu, Fuzhou ^{[1
]}

Sun, Huiling ^{[1
]}

机构：

[1] Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing Key Lab Prevent & Control Infect Dis Live, Beijing, Peoples R China

来源：

PLOS ONE | 2016年 / 11卷 / 12期

基金：

中国国家自然科学基金;

关键词：

POLYMERASE-CHAIN-REACTION; 16S RIBOSOMAL-RNA; RT-PCR; HOUSEKEEPING GENES; INTERNAL CONTROL; EXPRESSION; NORMALIZATION; PASTEURELLA; TOOL; IDENTIFICATION;

D O I：

10.1371/journal.pone.0167736

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Real-time quantitative reverse transcription PCR (qRT-PCR) offers a robust method for measurement of gene expression levels. Selection of reliable reference gene(s) for gene expression study is conducive to reduce variations derived from different amounts of RNA and cDNA, the efficiency of the reverse transcriptase or polymerase enzymes. Until now reference genes identified for other members of the family Pasteurellaceae have not been validated for Avibacterium paragallinarum. The aim of this study was to validate nine reference genes of serovars A, B, and C strains of A. paragallinarum in different growth phase by qRT-PCR. Three of the most widely used statistical algorithms, geNorm, NormFinder and Delta CT method were used to evaluate the expression stability of reference genes. Data analyzed by overall rankings showed that in exponential and stationary phase of serovar A, the most stable reference genes were gyrA and atpD respectively; in exponential and stationary phase of serovar B, the most stable reference genes were atpD and recN respectively; in exponential and stationary phase of serovar C, the most stable reference genes were rpoB and recN respectively. This study provides recommendations for stable endogenous control genes for use in further studies involving measurement of gene expression levels.

引用

页数：14

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