斑点叉尾鮰IL-22基因克隆及对不同刺激物的响应
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现代农业产业技术体系(CARS-49);盐城渔业高质量发展项目(YCSCYJ2021026)


Cloning of channel catfish (Ictalurus punctatus) interleukin-22 gene and its response to different stimuli
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    摘要:

    为探究白细胞介素-22(interleukin-22, IL-22)基因编码序列(coding sequence, CDS)的分子特征以及在不同病原微生物胁迫下IL-22的响应情况,以斑点叉尾鮰为(Ictalurus punctatus)研究对象,采用PCR扩增技术对该序列进行克隆,通过实时荧光定量PCR(qRT-PCR)方法分析各组织分布以及不同诱导模式下IL-22 mRNA表达量的变化情况。结果显示:该序列CDS 长度为534 bp,编码177个氨基酸;氨基酸序列多重比对分析发现IL-22在鱼类进化中与脊椎动物保守性较高。qRT-PCR结果显示,IL-22广泛分布在斑点叉尾鮰的10个组织中,表达量在后肠中最高,心脏中最低。Poly(I:C)、LPS、PHA、PMA刺激斑点叉尾鮰肾细胞24~48 h后,Poly(I:C)和LPS刺激下IL-22呈上调反应,PHA、PMA刺激下IL-22呈下调反应;细菌、病毒注射实验以及溴氰菊酯浸泡鱼体的攻毒实验结果显示,鳃、皮肤、后肠、脾脏、中肾和头肾组织中IL-22表达量整体上调。上述结果表明,不同刺激下不同组织中IL-22表达模式具差异性,该基因可能参与黏膜和多种免疫反应且发挥关键作用。

    Abstract:

    In order to explore the molecular characteristics of the interleukin-22 (IL-22) gene in channel catfish (Ictalurus punctatus) and its response to different pathogenic microorganisms,the coding sequence (CDS) of IL?22 was cloned by PCR amplification,and the distribution of IL?22 mRNA in different tissues and under different stimulate were monitored by quantitative real-time PCR (qRT-PCR).The results showed that the CDS of IL?22 was 534 bp in length and encoded 177 amino acids.Multiple alignment and analysis of amino acid sequence showed that IL?22 was highly conserved in evolution of fish and vertebrates.qRT-PCR results showed that IL?22 was widely distributed in 10 tissues of channel catfish,with the highest expression in the hindgut and the lowest in the heart.After stimulation for 24-48 h,IL?22 was up-regulated by Poly(I:C) and LPS stimulation,and down-regulated by PHA and PMA stimulation in the channel catfish kidney cells.The bacteria,virus injection and cypermethrin immersion experiments showed that the expression of IL?22 in the gill,skin,hindgut,spleen,trunk kidney and head kidney was up-regulated overall.The above results showed that the expression patterns of IL?22 in different tissues under different stimulate are different,and this gene can participate and play a key role in mucosal and various immune responses.

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朱霞,杨移斌,张蕾,胥宁,艾晓辉.斑点叉尾鮰IL-22基因克隆及对不同刺激物的响应[J].华中农业大学学报,2022,41(3):211-220

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  • 收稿日期:2022-02-01
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  • 在线发布日期: 2022-06-15
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