Identification of reference genes in blood before and after entering the plateau for SYBR green RT-qPCR studies

PeerJ

article

Jun Xiao¹  Xiaowei Li¹  Juan Liu¹  Xiu Fan¹  Huifen Lei¹  Cuiying Li¹ 

[1] Department of Blood Transfusion, General Hospital of Air Force

关键词: Plateau;    Hypoxia;    Gene expression;    Expression stability;    Reference gene;   

DOI  :  10.7717/peerj.3726

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

BackgroundTibetans have lived at high altitudes for thousands of years, and they have unique physiological traits that enable them to tolerate this hypoxic environment. However, the genetic basis of these traits is still unknown. As a sensitive and highly efficient technique, RT-qPCR is widely used in gene expression analyses to provide insight into the molecular mechanisms underlying environmental changes. However, the quantitative analysis of gene expression in blood is limited by a shortage of stable reference genes for the normalization of mRNA levels. Thus, systematic approaches were used to identify potential reference genes.ResultsThe expression levels of eight candidate human reference genes (GAPDH, ACTB, 18S RNA, β2-MG, PPIA, RPL13A, TBP and SDHA) were assessed in blood from hypoxic environments. The expression stability of these selected reference genes was evaluated using the geNorm, NormFinder and BestKeeper programs. Interestingly, RPL13A was identified as the ideal reference gene for normalizing target gene expression in human blood before and after exposure to high-altitude conditions.ConclusionThese results indicate that different reference genes should be selected for the normalization of gene expression in blood from different environmental settings.

【 授权许可】

CC BY