Selection of a suitable reference gene for gene-expression studies in Trichomonas gallinae under various biotic and abiotic stress conditions

文献类型： 外文期刊

第一作者： Cai, Haiming

作者： Cai, Haiming;Zhu, Yibin;Liu, Yu;Liao, Shenquan;Li, Juan;Lv, Minna;Lin, Xuhui;Hu, Junjing;Song, Yongle;Chen, Xiangjie;Yin, Lijun;Zhang, Jianfei;Qi, Nanshan;Sun, Mingfei;Yan, Zhuanqiang;Fang, Siyun;Wang, Dingai;Shen, Hanqin

作者机构：

关键词： Trichomonas gallinae; Reference genes selection; Stability analysis; TgaAtg8 gene

期刊名称：GENE （ 影响因子：3.5； 五年影响因子：3.3 ）

ISSN： 0378-1119

年卷期： 2024 年 920 卷

页码：

收录情况： SCI

摘要： Trichomonas gallinae, a globally distributed protozoan parasite, significantly affects the pigeon -breeding industry. T. gallinae infection mainly causes yellow ulcerative nodules on the upper respiratory tract and crop mucosa of pigeons, impeding normal breathing and feeding and ultimately causing death. Real-time quantitative PCR (qPCR) is a crucial technique for gene -expression analysis in molecular biology. Reference -gene selection for normalization is critical for ensuring this technique's accuracy. However, no systematic screening or validation of T. gallinae reference genes has been reported. This study quantified the transcript levels of ten candidate reference genes in T. gallinae isolates with different genotypes and culture conditions using qPCR. Using the geNorm, NormFinder, and BestKeeper algorithms, we assessed these reference genes' stabilities and ranked them using RankAggreg analysis. The most stable reference gene was tubulin beta chain (TUBB), while the widely used reference genes TUBG and GAPDH demonstrated poor stability. Additionally, we evaluated these candidate reference genes' stabilities using the T. gallinae TgaAtg8 gene. On using TUBB as a reference gene, TgaAtg8 ' s expression profiles in T. gallinae isolates with different genotypes remained relatively consistent under various culture conditions. Conversely, using ACTB as a reference gene distorted the data. These findings provide valuable reference -gene -selection guidance for functional gene research and gene -expression analysis in T. gallinae.

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