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干扰素(interferons,IFNs)是一类能够诱导细胞进入抗病毒状态的分泌蛋白,是脊椎动物抵抗病毒入侵的第一道屏障。本实验以青鳉(Oryzias latipes)为实验材料,通过polyⅠ:C诱导处理,克隆得到一个Ⅰ型干扰素c DNA序列,命名为Ol IFNβ。Ol IFNβ基因含有4个外显子和3个内含子,具有典型的Ⅰ型干扰素的结构特点,如含有2个半胱氨酸残基和m RNA非稳定序列。氨基酸序列比对分析显示,Ol IFNβ与三刺鱼IFN1同源性最高,达54%。系统进化树分析表明,Ol IFNβ与三刺鱼干扰素的亲缘关系最近,且和其它鱼类Ⅰ型干扰素聚成一个分支。荧光定量PCR分析表明,Ol IFNβ表达具有时间和组织特异性。PolyⅠ:C诱导处理后,随着处理时间的延长Ol IFNβ表达量逐渐上调,6 h后达到最高,随后逐渐下调。其中脾脏变化最为显著,鳃和肠次之。本研究显示,Ol IFNβ在青鳉先天免疫应答过程中起重要作用,为将来进一步探索Ol IFNβ的功能提供了帮助。
Interferons (IFNs) are secreted proteins that induce cells to enter an antiviral state and are the first barrier to vertebrate resistance to virus invasion. In this experiment, Oryzias latipes was used as experimental material, and a type I interferon cDNA was cloned by poly I: C induction and named as Ol IFNβ. The Ol IFNβ gene contains four exons and three introns, and has the structural characteristics of typical type I interferons, such as containing two cysteine residues and m RNA non-stable sequences. Amino acid sequence alignment analysis showed that Ol IFNβ had the highest homology with tricorn fish IFN1, reaching 54%. Phylogenetic tree analysis showed that Ol IFNβ has the closest genetic relationship with the tricornitis interferon and forms a branch with other fish type I interferons. Fluorescent quantitative PCR analysis showed that the expression of Ol IFNβ was time-and tissue-specific. Poly I: C induction treatment, with the extension of treatment time Ol IFNβ expression gradually increased after 6 h and then reached the highest, and then gradually decreased. The spleen changes the most significant, followed by gills and gut. This study shows that Ol IFNβ plays an important role in the process of innate immune responses to barley, which will help to further explore the function of Ol IFNβ in the future.