家庭管理基因的稳定性和适用性在弗勒博托明沙中
Fabiana Sassù1, Barbora Vomáčková Kykalová1, Cecilia Stahl Vieira1
1Department of Parasitology, Faculty of Science, Charles University, Viničná 7, Prague, 128 00, Czech Republic.
Scientific reports
|October 7, 2024
概括
我们在沙中确定了定量PCR (qPCR) 最稳定的参考基因,这些基因是莱什曼病的重要载体. 推使用延长因子1-α (EF1-α) 和核糖体蛋白L32 (RiboL32) 来对这些疾病载体进行精确的基因表达研究.
科学领域:
- 载体生物学 载体生物学
- 分子昆虫学分子昆虫学
- 寄生虫学的寄生虫学
背景情况:
- 沙 (Lutzomyia和Phlebotomus) 传播莱什曼病,这是一个重大的全球健康问题.
- 对这些载体进行准确的基因表达分析对于了解疾病传播和制定控制策略至关重要.
- 定量PCR (qPCR) 依赖于稳定的内源控制基因以获得可靠的结果,但适合沙的基因尚未得到充分确立.
研究的目的:
- 为了评估Lutzomyia longipalpis和Phlebotomus papatasi沙中常用的内源控制基因的表达稳定性.
- 在各种实验条件中确定qPCR最可靠的参考基因,包括不同的发育阶段,性别,饮食,dsRNA注射和Leishmania感染.
- 为了评估基因表达研究的基础对的跨物种适用性,在相关的沙物种.
主要方法:
- 量化PCR (qPCR) 用于分析六个候选参考基因的表达:actin,α-tubulin,GAPDH,60S核糖体蛋白L8 (RiboL8) 和L32 (RiboL32),以及延长因子1-α (EF1-α).
- 对Lutzomyia longipalpis和Phlebotomus papatasi的基因表达稳定性进行了评估,涉及不同生命阶段 (幼虫,成年人),性别,饮食和实验性治疗 (dsRNA注射,Leishmania感染).
- 在多种沙物种中测试了初级配对的有效性,包括Lutzomyia migonei,Phlebotomus duboscqi,Phlebotomus perniciosus和Sergentomyia schwetzi.
主要成果:
- 延长因子1-α (EF1-α) 在各种条件下在L. longipalpis和P. papatasi中都表现出高表达稳定性.
- 核糖体蛋白L8 (RiboL8) 和L32 (RiboL32) 在特定环境中也表现出良好的稳定性 (例如,L. longipalpis幼虫,P. papatasi成年人,切割的组织,感染的肠道).
- 几对原料对表现出跨物种的实用性,使得在更广泛的沙媒介中进行基因表达分析.
结论:
- 建议EF1-α,RiboL8和RiboL32作为Lutzomyia和Phlebotomus沙中qPCR的可靠内源控制.
- 识别的参考基因和验证的原料对将提高莱什曼病载体中基因表达研究的准确性和标准化.
- 这项研究为推进沙生物学的分子研究和莱什曼病控制工作提供了必不可少的工具.
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