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LvCTL1与WSSV复制的相互关系研究

邱炜 陈义烘

邱炜, 陈义烘. LvCTL1与WSSV复制的相互关系研究[J]. 华南师范大学学报(自然科学版), 2020, 52(6): 52-56. doi: 10.6054/j.jscnun.2020094
引用本文: 邱炜, 陈义烘. LvCTL1与WSSV复制的相互关系研究[J]. 华南师范大学学报(自然科学版), 2020, 52(6): 52-56. doi: 10.6054/j.jscnun.2020094
QIU Wei, CHEN Yihong. The Relationship between LvCTL1 and WSSV Replication[J]. Journal of South China normal University (Natural Science Edition), 2020, 52(6): 52-56. doi: 10.6054/j.jscnun.2020094
Citation: QIU Wei, CHEN Yihong. The Relationship between LvCTL1 and WSSV Replication[J]. Journal of South China normal University (Natural Science Edition), 2020, 52(6): 52-56. doi: 10.6054/j.jscnun.2020094

LvCTL1与WSSV复制的相互关系研究

doi: 10.6054/j.jscnun.2020094
基金项目: 

国家自然科学基金项目 31960206

国家自然科学基金项目 1772895

贵州省自然科学基金项目 J-[2020]1Y334

详细信息
    通讯作者:

    陈义烘,教授,Email: 20181024@m.scnu.edu.cn

  • 中图分类号: TN27

The Relationship between LvCTL1 and WSSV Replication

  • 摘要: C型凝集素(CTL)是一类重要的先天性免疫因子,在对虾机体的免疫防御中具有重要的作用.该研究利用相对定量PCR实验检测基因表达量、绝对定量PCR实验检测病毒拷贝数、组织切片分析实验检测感染病毒细胞数.实验数据显示:WSSV感染能显著上调LvCTL1的表达;WSSV感染的LvCTL1敲降的凡纳滨对虾中,膜囊蛋白VP28表达量、病毒拷贝数和胃上皮细胞中的感染细胞数都显著高于注射dsEGFP对照组.研究结果表明:WSSV感染后,机体通过上调LvCTL1表达来抑制WSSV复制.
  • 图  1  虾鳃组织中LvCTL1表达水平的检测

    注:以LvEF-1α为内参; 0 h的表达量设为1;显著性差异用短线上标记表示,*代表P < 0.05,* *代表P < 0.01,a与b表示差异有统计学意义,P < 0.01.

    Figure  1.  The detection of the expression levels of LvCTL1 in gill of prawn

    图  2  经dsLvCTL1处理并感染WSSV的虾体内病毒感染水平检测

    注:VP28表达水平的检测以LvEF-1α为内参; PBS+WSSV组在24 h的VP28表达量设为1;*表示差异具有统计学意义(P < 0.05).

    Figure  2.  The detection of the virus infection level in the prawn silenced by dsLvCTL1 after WSSV challenge

    图  3  感染WSSV对虾胃上皮细胞的组织病理分析

    注:箭头指示含病毒包涵体的受感染细胞.

    Figure  3.  The histological pathology in stomach epithelium of the L.vannamei challenged by WSSV

    图  4  LvNF-κB对LvCTL1表达水平的影响

    注:以LvEF-1α为内参; 0 h (未经任何处理样本)表达水平设为1;*表示差异具有统计学意义(*P < 0.05; * *P < 0.01).

    Figure  4.  The effect of LvNF-κB on the expression of LvCTL1

    表  1  所用引物序列

    Table  1.   The primer sequences

    用途 引物 序列(5′ to 3′)
    dsRNA的合成 LvCTL1-F(T7) TAATACGACTCACTATAGGATGAAGTTCCTAGCGCCG
    LvCTL1-R(T7) TAATACGACTCACTATAGGTCAATATTTCTTGAGGCAAATG
    LvCTL1-F ATGAAGTTCCTAGCGCCG
    LvCTL1-R TCAATATTTCTTGAGGCAAATG
    EGFP-F(T7) TAATACGACTCACTATAGGTGCTTCAGCCGCTACCC
    EGFP-R(T7) TAATACGACTCACTATAGGCGCTTCTCGTTGGGGTC
    EGFP-F TGCTTCAGCCGCTACCC
    EGFP-R CGCTTCTCGTTGGGGTC
    RT-PCR实验 LvCTL1-F GGACGCTTATGTCACCTGC
    LvCTL1-R TGTGTGACATCCTTGCTCTTG
    LvEF-1α-F GTATTGGAACAGTGCCCGTG
    LvEF-1α-R ACCAGGGACAGCCTCAGTAAG
    VP28-F AACACCTCCTCCTTCACCC
    VP28-R GGTCTCAGTGCCAGAGTAGGT
    WSSV32678-F TGTTTTCTGTATGTAATGCGTGTAGGT
    WSSV32753-R CCCACTCCATGGCCTTCA
    TaqMan probe-WSSV32706 CAAGTACCCAGGCCCAGTGTCATACGTT
    下载: 导出CSV
  • [1] ASCHE F, ANDERSON J L, BOTTA R, et al. The economics of shrimp disease[J]. Journal of Invertebrate Pathology, 2020:107397/1-9.
    [2] LI C Z, WANG S, HE J G. The two NF-κB pathways regulating bacterial and WSSV infection of shrimp[J]. Frontiers in Immunology, 2019, 10:1785/1-26.
    [3] SANCHEZ-PAZ A. White spot syndrome virus:an overview on an emergent concern[J]. Veterinary Research, 2010, 41(6):43/1-34.
    [4] SONG F, CHEN G L, LU K C, et al. Identification and functional characterization of a C-type lectin gene from Litopenaeus vannamei that is associated with ER-stress response[J]. Fish and Shellfish Immunology, 2019, 41:977-985.
    [5] WANG X W, WANG J X. Crustacean hemolymph microbiota:endemic, tightly controlled, and utilization expectable[J]. Molecular Immunology, 2015, 68(2):404-411.
    [6] HUANG Y, REN Q. Research progress in innate immunity of freshwater crustaceans[J]. Developmental and Comparative Immunology, 2020, 104:103569/1-9.
    [7] ZHAO Z Y, YIN Z X, XU X P, et al. A novel C-type lectin from the shrimp Litopenaeus vannamei possesses anti-white spot syndrome virus activity[J]. Journal of Virology, 2009, 41(1):347-356.
    [8] QIU W, ZHANG S, CHEN Y G, et al. Litopenaeus vannamei NF-κB is required for WSSV replication[J]. Developmental and Comparative Immunology, 2014, 45(1):156-162.
    [9] PFAFFL M W. A new mathematical model for relative quantification in real-time RT-PCR[J]. Nucleic Acids Research. 2001, 29(9):e45/1-6.
    [10] ROBALINO J, BROWDY C L, PRIOR S, et al. Induction of antiviral immunity by double-stranded RNA in a marine invertebrate[J]. Journal of Virology, 2004, 78(19):10442-10448.
    [11] WANG Y G, HASSAN M D, SHARIFF M, et al. Histopathology and cytopathology of white spot syndrome virus(WSSV) in cultured Penaeus monodon from peninsular Malaysia with emphasis on pathogenesis and the mechanism of white spot formation[J]. Diseases of Aquatic Organisms, 1999, 39(1):1-11.
    [12] VAN HULTEN M C, WITTEVELDT J, SNIPPE M, et al. White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp[J]. Virology, 2001, 285(2):228-233.
    [13] WITTEVELDT J, CIFUENTES C C, VLAK J M, et al. Protection of Penaeus monodon against white spot syndrome virus by oral vaccination[J]. Journal of Virology, 2004, 78(4):2057-2061.
    [14] WAN F, LENARDO M J. The nuclear signaling of NF-kappa B:current knowledge, new insights, and future perspectives[J]. Cell Research, 2010, 20(1):24-33.
    [15] OECKINGHAUS A, HAYDEN M S, GHOSH S. Crosstalk in NF-κB signaling pathways[J]. Nature Immunology, 2011, 12(8):695-708.
    [16] SURABHI R M, GAYNOR R B. RNA interference directed against viral and cellular targets inhibits human immunodeficiency Virus Type 1 replication[J]. Journal of Virology, 2002, 76(24):12963-12973.
    [17] WEST M J, LOWE A D, KARN J. Activation of human immunodeficiency virus transcription in T cells revisited:NF-kappa B p65 stimulates transcriptional elongation[J]. Journal of Virology, 2001, 75(18):8524-8537.
    [18] SAKAKIBARA S, SAKAKIBARA K, TOSATO G. NF-kappa B activation stimulates transcription and replication of retrovirus XMRV in human B-lineage and prostate carcinoma cells[J]. Journal of Virology, 2011, 85(7):3179-3186.
    [19] HUANG X D, YIN Z X, LIAO J X, et al. Identification and functional study of a shrimp Relish homologue[J]. Fish and Shellfish Immunolog, 2009, 27(2):230-238.
    [20] HUANG X D, YIN Z X, JIA X T, et al. Identification and functional study of a shrimp dorsal homologue[J]. Developmental and Comparative Immunology, 2010, 34(2):107-113.
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出版历程
  • 收稿日期:  2020-08-24
  • 刊出日期:  2020-12-25

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