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在各种生物和非生物压力条件下选择合适的基因参考基因用于Trichomonas gallinae基因表达研究

Haiming Cai¹, Yibin Zhu¹, Yu Liu¹

¹Key Laboratory of Livestock Disease Prevention of Guangdong Province, Key Laboratory of Avian Influenza and Other Major Poultry Diseases Prevention and Control, Ministry of Agriculture and Rural Affairs, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.

|May 4, 2024

概括

在Trichomonas gallinae中确定qPCR的最佳基因对准确的基因表达分析至关重要. 图布林β链 (TUBB) 证明最稳定,表现优于常用的基因,如GAPDH.

科学领域:

寄生虫学的寄生虫学
分子生物学分子生物学
生物信息学是一种生物信息学.

背景情况:

类是一种重要的原生虫寄生虫,影响着全球子繁殖产业.
感染导致性病变,导致呼吸和食问题,以及子的死亡率.
使用定量PCR (qPCR) 精确的基因表达分析依赖于稳定的参考基因进行正常化.

研究的目的:

系统地选和验证Trichomonas gallinae中的qPCR候选参考基因.
在T. gallinae.中确定最稳定的基因参考基因以进行可靠的基因表达研究.

主要方法:

在不同基因型和培养条件的T. gallinae分离物中,使用qPCR量化了10个候选参考基因.
基因表达稳定性使用geNorm,NormFinder和BestKeeper算法进行评估.
排名稳定性通过RankAggreg分析来确定.

主要成果:

结核素β链 (TUBB) 被确定为最稳定的参考基因.
广泛使用的参考基因TUBG和GAPDH表现出不良的稳定性.
使用TUBB来规范TgaAtg8表达产生了一致的结果,而ACTB则扭曲了数据.

结论:

TUBB被推作为Trichomonas gallinae中qPCR的最佳参考基因.
这项研究为T. gallinae功能基因组学研究中选择参考基因提供了必要的指导.
准确的正常化对于在这个经济上重要的寄生虫中可靠的基因表达分析至关重要.

关键词:

基因参考基因的选择选择.稳定性分析是一种稳定性分析.在TgaAtg8基因基因.这种植物是Trichomonas gallinae.

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