团头鲂补体因子Bf/C2的克隆和表达分析
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中央高校基本科研业务费专项(2662015PY134); 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)


Cloning and expression of complement factor B and C2 (Bf/C2) gene in blunt snout bream(Megalobrama amblycephala)
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    摘要:

    为探索团头鲂(Megalobrama amblycephala)补体因子Bf和C2(complement factor B/C2,Bf/C2)的可能功能,在转录组数据基础上,采用RT-PCR克隆得到Bf/C2A和Bf/C2B基因的cDNA序列;采用荧光实时定量PCR技术检测了两基因在团头鲂早期发育过程、健康成鱼及感染嗜水气单胞菌后各组织中的表达变化。结果显示,Bf/C2A cDNA全长2 520 bp,包含5′ UTR 42 bp、ORF 2 298 bp、3′ UTR 180 bp,编码765个氨基酸。Bf/C2B基因ORF全长2 130 bp,编码710个氨基酸。两基因的氨基酸序列分别与鲤B/C2 A2及草鱼Bf/C2B相似性最高。在早期发育阶段Bf/C2A和Bf/C2B均在在出膜后1 d表达量最高,肠管形成期次之,其他时期的表达量相对较低;两基因在健康成鱼10个组织中均有表达,肝脏中表达量最高,肾脏、头肾、脾脏次之,其他组织表达量相对较低;在嗜水气单胞菌感染后,两基因在免疫相关组织如肝脏和脾脏中的表达均显著上升。上述结果表明Bf/C2在应对细菌感染的免疫过程中发挥着重要的作用。

    Abstract:

    To explore the possible function of the complement factor B and C2 (Bf/C2) gene in blunt snout bream (Megalobrama amblycephala), the cDNA sequences were cloned with RT-PCR and the expression of the Bf/C2mRNA in healthy fish tissues and after Aeromonas hydrophila infection during early development was investigated with quantitative realtime PCR.The results showed that the sequence length of Bf/C2A cDNA was 2 520 bp containing a 42 bp 5′ UTR,a 2 298 bp ORF and a 180 bp 3′ UTR,encoding 765 amino acid.The Bf/C2B cDNA had an 2 131 bp ORF,encoding 710 amino acid.The mRNA of both Bf/C2A and Bf/C2B were highly expressed at 1 dpf,followed by the intestinal formation stage during early development.The highest expression of both Bf/C2A and Bf/C2B in healthy adults appeared in the liver,followed by the kidney,head kidney and spleen.Theexpression of Bf/C2A and Bf/C2B exhibited a significant change after infection of A. hydrophila in the inmmunityrelated tissues including the liver and kidney,indicating that Bf/C2 plays important role in the immune response against bacterial infection.

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范君,张锋,王卫民,刘红.团头鲂补体因子Bf/C2的克隆和表达分析[J].华中农业大学学报,2019,38(2):30-37

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  • 收稿日期:2018-09-27
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  • 在线发布日期: 2019-01-30
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