中华鲟IFN-γ的免疫调控作用
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S941

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农业农村部淡水生物多样性保护重点实验室开放基金(LFBC 1007)


Immune regulation role of IFN-γ in Acipenser sinensis
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    摘要:

    为了解中华鲟IFN-γ的免疫调控作用,实验通过中华鲟转录组获得IFN-γ cDNA序列,构建重组表达质粒pTRI-st-IFNγ,原核表达IFN-γ蛋白,检测其在病毒和细菌感染过程中的免疫调控作用。SDS-PAGE显示,中华鲟IFN-γ重组蛋白大小为19.17 ku,以可溶性表达为主,浓度为0.998 mg/mL;荧光定量PCR显示,IFN-γ蛋白能够诱导中华鲟肾脏细胞中下游相关抗病毒基因MxViperinCXCL家族中CXCL11-L1、CXCL11-L2表达;与EPC细胞共孵育能够有效抑制鲤春病毒血症病毒(spring viraemia of carp virus,SVCV)引起的细胞病变和增殖,并对SVCV病毒的GPN 3个基因表达量呈现出极显著性下调;在体外能够调控抑制嗜水气单胞菌、中间产气单胞菌和维氏气单胞菌并对其生长表现出明显的抑制作用。

    Abstract:

    Acipenser sinensis belongs to the cartilaginous fish, which is between the cartilaginous fishes and teleost fishes. It is an important branch point in the evolutionary tree of the biological immune system. It is a living fossil for the study of the evolution of fish and vertebrates, and plays an extremely important role in the evolutionary history of fish and vertebrates. In order to understand the immunomodulatory effect of A. sinensis IFN-γ, IFN- γ sequence was obtained by transcriptome of A. sinensis, the recombinant plasmid of pTRI-st-IFNg was constructed and transformed into the BL21 Escherichia coli for expression and expressed in competent prokaryotic cells. SDS-PAGE analysis showed that the size of IFN-γ recombinant protein was 19.17 ku, and the concentration was 0.998 mg/mL. Quantitative PCR showed that the antiviral genes Mx, Viperin, CXCL11-L1,CXCL11-L2 (CXC subfamily of chemokines) and antiviral protein Epithelioma papulosum cyprini cells (EPC) P,N,G were induced, which indicated that IFN-λ activated antiviral activity. Additionally, spring viremia of carp virus (SVCV) replication as well as the production of cytopathic effect (CPE) was significantly inhibited. Results of incubation with Aeromonas hydrophila, A. intermedia and A. veronii showed a significant inhibition on their growth and activity. In conclusion, this study provides partial theory for the understanding of IFN-γ bioactivity in A. sinensis.

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郑楚文,袁汉文,田光明,李骞,张书环,许巧情,危起伟.中华鲟IFN-γ的免疫调控作用[J].水产学报,2020,44(9):1539~1548

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  • 收稿日期:2020-07-29
  • 最后修改日期:2020-08-25
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  • 在线发布日期: 2020-09-27
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