团头鲂TYK2基因的克隆与表达分析
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国家自然科学基金项目(31972781);现代农业产业技术体系国家大宗淡水鱼产业技术体系建设专项(CARS-46-08)


Molecular cloning and expression of TYK2 in Megalobrama amblycephala
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    摘要:

    以团头鲂为研究对象,克隆获得络氨酸激酶2(tyrosine kinase 2,TYK2)基因的ORF序列,该序列长3 489 bp,编码1 162 aa。团头鲂TYK2基因包含23个外显子和22个内含子,与大多数脊椎动物的基因结构相似。蛋白质结构域预测结果显示,TYK2包含4个结构域:B41、SH2、TyrKc(假激酶区)和TyrKc(络氨酸激酶区)。氨基酸多序列比对和系统进化分析结果均显示,TYK2在进化过程中非常保守。荧光定量PCR结果显示,在健康成鱼中,TYK2在中肾的表达量最高,其次是血液、脾脏和头肾,在肠道、脑、鳃、心脏、肌肉和肝脏中的表达量较低。感染嗜水气单胞菌后,TYK2在脾脏和肠中有相似的表达模式:在感染后4 h显著降低并达到最小值,在12 h显著上升且达到最大值(P<0.05);在中肾中,在24 h极显著上升且到达最大值,在36 h极显著降低并达到最小值。以上研究结果表明,TYK2基因在团头鲂抗嗜水气单胞菌感染的过程中发挥着重要的作用。

    Abstract:

    Jak kinase is a non-receptor type tyrosine kinase,which is broadly found in mammals,birds,insects and fish. It is an important factor in Janus kinase/signal transduction and transcription factor signaling pathway,which is widely involved in the processes of immunity,cell proliferation,growth,differentiation,migration and apoptosis. TYK2 is the first member of the Jak family to be identified and has been confirmed to play a key role in cytokine transduction. In this article,the ORF sequence of the TYK2 gene in Megalobrama amblycephala was cloned and analyzed. Results showed that the ORF sequence of the TYK2 gene in M. amblycephalawas 3 489 bp in length encoding 1 162 aa. The M. amblycephala TYK2 (MamTYK2) is consisted of 23 exons and 22 introns,similar to other vertebrate species. The MamTYK2 protein is composed of 4 domains including B41,SH2,TyrKc (pseudokinase domain) and TyrKc (tyrosine kinase domain). TYK2 is highly conserved through evolution. Based on the results of quantitative realtime PCR (qRT-PCR),among healthy adult tissues,TYK2 is highly expressed in the middle kidney,followed by the blood,spleen,and head kidney,and lowly expressed in other tissues including the intestine,brain,gill,heart,muscle,and liver. After infected by Aeromonas hydrophila,the TYK2 mRNA showed a similar expression pattern in the spleen and intestine:significantly decreased at 4 hpi (hour post infection) (P<0.05) and reached a minimum,and significantly increased at 12 hpi (P<0.05) and reached a maximum. It significantly increased and reached the maximum value at 24 hpi (P<0.01),and significantly decreased and reached the minimum value at 36 hpi (P<0.01) in the midkidney. It is indicated that the MamTYK2 gene may play an important role in the process of M. amblycephala resistance against A. hydrophila.

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张建,王济秀,吴晓丽,刘红.团头鲂TYK2基因的克隆与表达分析[J].华中农业大学学报,2021,40(2):197-205

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  • 收稿日期:2020-05-03
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  • 在线发布日期: 2021-04-16
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