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
作者： Cai, Haiming 1 ; Zhu, Yibin 1 ; Liu, Yu 1 ; Yan, Zhuanqiang 2 ; Shen, Hanqin 3 ; Fang, Siyun 2 ; Wang, Dingai 2 ; Liao, Shenquan 1 ; Li, Juan 1 ; Lv, Minna 1 ; Lin, Xuhui 1 ; Hu, Junjing 1 ; Song, Yongle 1 ; Chen, Xiangjie 1 ; Yin, Lijun 1 ; Zhang, Jianfei 1 ; Qi, Nanshan 1 ; Sun, Mingfei 1 ;
作者机构： 1.Guangdong Acad Agr Sci, Key Lab Livestock Dis Prevent Guangdong Prov, Key Lab Avian Influenza & Other Major Poultry Dis, Inst Anim Hlth,Minist Agr & Rural Affairs, Guangzhou 510640, Peoples R China
2.Wens Foodstuffs Grp Co Ltd, Wens Grp Acad, Xinxing 527400, Guangdong, Peoples R China
3.Guangdong Jingjie Inspection & Testing Co Ltd, Xinxing 527400, Guangdong, Peoples R China
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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