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七彩神仙鱼C型凝集素家族成员CD302和MBL的克隆及在病原刺激下的免疫应答

马腾飞 温彬 刘鑫 缪琳 高建忠 陈再忠

马腾飞, 温彬, 刘鑫, 缪琳, 高建忠, 陈再忠. 七彩神仙鱼C型凝集素家族成员CD302和MBL的克隆及在病原刺激下的免疫应答[J]. 南方水产科学. doi: 10.12131/20220018
引用本文: 马腾飞, 温彬, 刘鑫, 缪琳, 高建忠, 陈再忠. 七彩神仙鱼C型凝集素家族成员CD302和MBL的克隆及在病原刺激下的免疫应答[J]. 南方水产科学. doi: 10.12131/20220018
MA Tengfei, WEN Bin, LIU Xin, MIAO Lin, GAO Jianzhong, CHEN Zaizhong. Molecular cloning and characterization in immune response of two C-type lectin genes (SaCD302 and SaMBL) from Symphysodon aequifasciatus[J]. South China Fisheries Science. doi: 10.12131/20220018
Citation: MA Tengfei, WEN Bin, LIU Xin, MIAO Lin, GAO Jianzhong, CHEN Zaizhong. Molecular cloning and characterization in immune response of two C-type lectin genes (SaCD302 and SaMBL) from Symphysodon aequifasciatus[J]. South China Fisheries Science. doi: 10.12131/20220018

七彩神仙鱼C型凝集素家族成员CD302和MBL的克隆及在病原刺激下的免疫应答

doi: 10.12131/20220018
基金项目: 上海市自然科学基金项目  (20ZR1423600);上海扬帆人才计划项目 (19YF1419400)
详细信息
    作者简介:

    马腾飞 (1998—),男,硕士研究生,研究方向为观赏鱼养殖。E-mail: tfmalh@163.com

    通讯作者:

    陈再忠 (1972—),男,教授,博士,从事观赏鱼养殖研究。E-mail : chenzz@shou.edu.cn

  • 中图分类号: S 917.4

Molecular cloning and characterization in immune response of two C-type lectin genes (SaCD302 and SaMBL) from Symphysodon aequifasciatus

  • 摘要: 七彩神仙鱼 (Symphysodon aequifasciatus) 具独特的亲代抚育行为,前期研究发现2种C型凝集素在育幼亲本皮肤高表达,但表达模式不同,预示着潜在的功能分化。为探究两者潜在免疫功能的差异性,克隆获得了七彩神仙鱼C型凝集素基因SaCD302和SaMBL,并分析了病原刺激下的免疫应答模式。结果显示,SaCD302和SaMBL的ORF区长度分别为741 bp和795 bp,分别编码246和264个氨基酸。SaCD302序列包含一个信号肽 (aa 1~29),一个CTL结构域 (aa 32~169),一个跨膜结构域 (aa 184~206)。SaMBL序列包含两个低复杂度区域 (aa 32~92和aa 104~121) 和一个典型的CTL结构域 (aa 137~263)。SaCD302和SaMBL的氨基酸序列与尼加拉瓜湖始丽鱼 (Archocentrus centrarchus) 对应基因序列的同源性最高。SaCD302在各组织均表达,头肾表达量最高,其次为肝脏和鳃;SaMBL在皮肤高表达,在肠道、肝脏、脾脏和性腺低表达。嗜水气单胞菌 (Aeromonas hydrophila) 感染后,SaCD302在脾脏、头肾、肠道和皮肤均显著上调,SaMBL仅在头肾、肠道和皮肤中表达上调,且时序性不同,表明二者可能在七彩神仙鱼免疫防御中发挥不同作用。
  • 图  1  SaCD302 (a)和SaMBL (b)基因的核酸序列及氨基酸序列

    Figure  1.  Nucleotide and deduced amino acid sequence of SaCD302 (a) and SaMBL (b)

    图  2  SaCD302(a) 和SaMBL (b) 结构域示意图

    Figure  2.  Domain of SaCD302(a) and SaMBL(b)

    图  3  SaCD302 (a) 和SaMBL (b) 氨基酸的多重序列比对

    Figure  3.  Multiple sequence alignment of amino acid sequences of SaCD302(a) and SaMBL (b)

    图  4  基于NJ法构建的SaCD302 (a) 和SaMBL (b) 系统进化树

    Figure  4.  Neighbor-joining phylogenetic analysis of SaCD302 (a) and SaMBL (b)

    图  5  SaCD302 (a) 和SaMBL (b) 在七彩神仙鱼不同组织中的表达特征

    Figure  5.  Relative expression of SaCD302(a) and SaMBL (b) in different tissues of S. aequifasciatus

    图  6  嗜水气单胞菌感染后SaCD302在七彩神仙鱼不同组织中的表达分析

    Figure  6.  Expression analysis of indifferent tissues of SaCD302 from S. aequifasciatus infected by A. hydrophila

    图  7  嗜水气单胞菌感染后SaMBL在七彩神仙鱼不同组织中的表达分析

    Figure  7.  Expression analysis of SaMBL in different tissues from S. aequifasciatus nfected by A. hydrophila i

    表  1  七彩神仙鱼SaCD302SaMBL基因克隆及荧光定量PCR引物

    Table  1.   Cloning and fluorescence quantitative primers of SaCD302 and SaMBL gene in S. aequifasciatus

    引物
    Primer
    引物序列5′—3′
    Primer sequence
    用途
    Function
    SaCD302-F ATGGAGTCGCGGAAGAAAAGT 基因克隆
    SaCD302-R CTATGGCACGCTGTCAGTTTC 基因克隆
    SaCD302-qPCR-F TGTGAAGCTGCTCAGGATGA 荧光定量PCR
    SaCD302-qPCR-R
    SaMBL-F
    SaMBL-R
    SaMBL-qPCR-F
    SaMBL-qPCR-R
    GCTTCTGGTGCAGAAACCAA
    ATGTGCATTGTTCTCTCGGA
    CTATAACTGACAAATGATGT
    GGTCACGGTACTTTCTGGGA
    AGCCTCTCTCCTTGTTGGAC
    荧光定量PCR
    基因克隆
    基因克隆
    荧光定量PCR
    荧光定量PCR
    β-actin-qPCR-F GTGCGTGACATCAAGGAGAAG 荧光定量PCR
    β-actin-qPCR-R GGAAGGAAGGCTGGAAGAGG 荧光定量PCR
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-01-17
  • 修回日期:  2022-03-31
  • 录用日期:  2022-05-07
  • 网络出版日期:  2022-05-13

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