柔嫩艾美耳球虫Etron2基因在不同发育阶段转录水平差异的分析

Transcription Profiles of Etron2 Gene during the Different Development Stages

为研究柔嫩艾美耳球虫不同发育时期Etron2基因的转录水平差异,选取E.tenella生活史中的未孢子化卵囊、孢子化卵囊、子孢子、第二代裂殖子,分别提取总RNA,反转录为cDNA;选择Etactin作为参照基因,Etron2为目的基因,设计实时荧光定量PCR的引物,分析在E.tenella生活史中不同发育阶段Etron2转录水平的差异。结果显示Etron2在未孢子化卵囊中转录水平最高,然后分别是裂殖子、孢子化卵囊和子孢子。在柔嫩艾美耳球虫发育过程中,Etron2基因可能在未孢子化卵囊阶段被转录储备,随着宿主细胞的刺激,得以大量表达,在入侵过程中发挥作用。

The objective of this study was to study the transcription profiles of Etron2 gene during the different development stages.The parasites at different developmental stages were harvested and their total RNA were substracted by real-time PCR using Etactin gene as a reference for establishing standard curves to evaluate the copy number of Etron2 genes.All of quantitative analyses were reliable because the melting curve showed a single peak.Etron2 gene transcription reached a peak during the unsporulated oocysts stage,then merozoites,sporulated oocysts and the lowest during the sporozoites stage.It is likely that the transcription messages were stored during the unsporulated oocysts stage,and then the Etron2 protein can be translated and secreted into the rhoptry soon when the oocysts were sporulated,and it may play an important role in the process of invasion.