脊尾白虾Strawberry Notch1基因的克隆及免疫功能

Cloning and immunological function of SBNO1 gene in Exopalaemon carinicauda

  • 摘要: 为探究Strawberry Notch1基因(SBNO1)在甲壳动物中的免疫功能,实验对脊尾白虾SBNO1的cDNA序列全长、系统进化、组织分布以及副溶血性弧菌感染后SBNO1和Notch信号通路中相关基因表征进行了研究。结果显示,脊尾白虾SBNO1 cDNA序列全长4 353 bp,共编码1 380个氨基酸,预测相对分子质量158.91 ku,理论等电点4.81。qPCR分析显示,脊尾白虾SBNO1在各组织中均有表达,其中在肝胰腺中表达量最高,性腺次之。利用RNAi干扰SBNO1显示,脊尾白虾在感染副溶血性弧菌后,干扰组死亡率显著高于未干扰组。副溶血性弧菌感染脊尾白虾后显著影响了Notch信号通路中SBNO1、NumbDelta在肝胰腺中的转录情况,而Jagged的表达差异不显著。SBNO1和Numb的表达量随时间延长都呈现出先上升后下降的趋势,Delta的表达量呈现出先下降后上升的趋势。研究表明,Notch信号通路参与了脊尾白虾副溶血性弧菌感染后的免疫。本研究有助于阐明无脊椎动物先天免疫的机制,为抗病脊尾白虾的分子育种提供理论依据。

     

    Abstract: In order to explore the immune function of Strawberry Notch1 gene (SBNO1) in crustaceans, the full length of SBNO1 gene cDNA sequence, phylogeny, tissue distribution, the expression of SBNO1 and related genes in Notch signaling pathways by Vibrio parahaemolyticus (VP) injection were studied in this paper. The results showed that the cDNA sequence of SBNO1 was 4 353 bp, encoding a total of 1380 amino acids. The predicted molecular weight of protein is 158.91 ku and theoretical isoelectric point is 4.8. According to the RT-qPCR analysis, it can be found that SBNO1 was expressed in all tested tissues, among which hepatopancreas had the highest level and gonad followed. Futhermore, after RNAi interference, compared with the non-inference group, the death rate of the inference group injected with VP was significantly higher. The transcription of SBNO1, Numb and Delta in the hepatopancreas in Notch signaling was obviously influenced by VP. However, there was no significant difference in the expression of Jagged. In the group injected with VP, the expression levels of SBNO1 and Numb increased first and then decreased over time, while the expression of Delta decreased first and then increased. The results indicate that Notch signaling pathway plays an important role in the immunity of E. carinicauda by VP injection. This study helps to clarify the mechanism of invertebrate innate immunity and provides a theoretical basis for molecular breeding of E. carinicauda.

     

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