Validation of Reference Genes for Quantitative Real-Time PCR in Bovine PBMCs Transformed and Non-transformed by Theileria annulata
文献类型: 外文期刊
第一作者: Zhao, Hongxi
作者: Zhao, Hongxi;Liu, Junlong;Li, Youquan;Yang, Congshan;Zhao, Shuaiyang;Liu, Juan;Liu, Aihong;Liu, Guangyuan;Yin, Hong;Guan, Guiquan;Luo, Jianxun;Yin, Hong;Zhao, Hongxi
作者机构:
关键词: Theileria annulata;quantitative real-time PCR (qPCR);reference gene;signaling pathway
期刊名称:KOREAN JOURNAL OF PARASITOLOGY ( 影响因子:1.341; 五年影响因子:1.669 )
ISSN: 0023-4001
年卷期: 2016 年 54 卷 1 期
页码:
收录情况: SCI
摘要: Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (beta-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.
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