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罗氏沼虾nanos1基因克隆、表达及亚细胞定位

周南 王亚坤 洪坤浩 魏捷 于凌云 朱新平

周南, 王亚坤, 洪坤浩, 魏捷, 于凌云, 朱新平. 罗氏沼虾nanos1基因克隆、表达及亚细胞定位[J]. 南方水产科学. doi: 10.12131/20220067
引用本文: 周南, 王亚坤, 洪坤浩, 魏捷, 于凌云, 朱新平. 罗氏沼虾nanos1基因克隆、表达及亚细胞定位[J]. 南方水产科学. doi: 10.12131/20220067
ZHOU Nan, WANG Yakun, HONG Kunhao, WEI Jie, YU Lingyun, ZHU Xinping. Molecular cloning, expression profile and subcellular localization of nanos1 gene (Macrobrachium rosenbergii)[J]. South China Fisheries Science. doi: 10.12131/20220067
Citation: ZHOU Nan, WANG Yakun, HONG Kunhao, WEI Jie, YU Lingyun, ZHU Xinping. Molecular cloning, expression profile and subcellular localization of nanos1 gene (Macrobrachium rosenbergii)[J]. South China Fisheries Science. doi: 10.12131/20220067

罗氏沼虾nanos1基因克隆、表达及亚细胞定位

doi: 10.12131/20220067
基金项目: 中国水产科学研究院珠江水产研究所所级基本科研业务费 (2022SJ-XT1);中国水产科学研究院 (珠江水产研究所) 科技创新团队 (2020TD35, 2020ZJTD01);广东省促进经济发展专项资金 (粤农2019B7);中国-东盟渔业资源保护与开发利用项目 (CAMC-2018F)
详细信息
    作者简介:

    周南:周 南 (1997—),女,硕士研究生,研究方向为淡水鱼类生殖发育学。E-mail: 932085618@qq.com

    通讯作者:

    于凌云 (1980—), 男,副研究员,博士,从事水产育种研究。E-mail: lysnp@163.com

    朱新平 (1964—),男,研究员,博士,从事水产育种研究。E-mail: zhuxinping_1964@163.com

  • 中图分类号: S 917.4

Molecular cloning, expression profile and subcellular localization of nanos1 gene (Macrobrachium rosenbergii)

  • 摘要: 为探究nanos1基因在罗氏沼虾 (Macrobrachium rosenbergii) 生殖发育中的功能,克隆了罗氏沼虾nanos1基因,其cDNA序列长2 811 bp,编码243个氨基酸;系统进化分析结果表明其与中华绒螯蟹 (Eriocheir sinensis) 的同源性最高,与鱼类的nanos1同源性较高,与哺乳类同源性较低。半定量PCR检测发现,nanos1在卵巢中特异表达;通过实时荧光定量PCR检测该基因在不同胚胎发育时期及幼体中的表达水平,结果显示:nanso1 mRNA在 (卵) 未受精时期表达量最高,显著高于 (卵) 受精期及胚胎发育各期 (P<0.05);在胚胎发育期间, (卵) 受精时期表达最高,显著高于卵裂期且极显著高于胚胎发育后期 (P<0.01);该基因在卵裂期的表达水平显著高于囊胚期至幼体期;而囊胚期至幼体期之间表达水平较低且无差异。原位杂交技术检测显示,nanos1 mRNA在卵原细胞及初级卵母细胞 (Oc1,Oc2,Oc3,Oc4) 的细胞质中表达。以上结果表明,nanos1与罗氏沼虾雌性生殖细胞发育有着密切的关系。
  • 图  1  罗氏沼虾Mrnanos1 cDNA序列及氨基酸序列

    Figure  1.  Sequence and amino acid sequence of Mrnanos1 cDNA from M. rosenbergii

    图  2  MrNanos1氨基酸序列与其他物种Nanos家族蛋白的系统进化树分析

    Figure  2.  Phylogenetic tree analysis of MrNanos1 amino acid sequence with Nanos family proteins of other species

    图  3  罗氏沼虾MrNanos1与其他物种锌指区域结构的比较

    Figure  3.  Multiple alignments analysis of zinc finger domain of MrNanos1 in M. rosenbergii compared with that of various species

    图  4  Mrnanos1基因在罗氏沼虾不同组织中的表达分析

    Figure  4.  Expression analysis of Mrnanos1 gene in different tissues of M. rosenbergii

    图  5  罗氏沼虾nanos1在不同胚胎发育时期及早期幼体中的相对表达量

    Figure  5.  Relative expression level of nanos1 in different embryonic develomental stages and larvae of M. rosenbergii

    图  6  Mrnanos1 mRNA 在罗氏沼虾卵巢中的亚细胞分布

    Figure  6.  Subcellular localization of Mrnanos1 mRNA in ovary of M. rosenbergii

    表  1  本研究所用的全部引物

    Table  1.   All primers used in this study

    引物
    Primer
    序列 (5'—3')
    Sequence (5'—3')
    长度
    Length/bp
    温度
    Temperature/ ℃
    应用
    Application
    nanos1 F
    nanos1 R
    AAAACAACTAACGGCGATGC
    ACGAAGACCCGCTGATATTG
    74757RT-PCR, PCR, ISH
    nanos1 QF
    nanos1 QR
    GGCCTTAGGGCAAATAGCTC
    ACGAAGACCCGCTGATATTG
    23360qPCR
    β-actin F
    β-actin R
    GAAGCGTACATGGTGGGACT
    GGTGACCTGCGAGATTCATT
    46657RT-PCR
    β-actin QF
    β-actin QR
    CAGGGAAAAGATGACCCAGA
    GGAAGTGCATACCCCTCGTA
    16160qPCR
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-03-15
  • 修回日期:  2022-04-18
  • 录用日期:  2022-05-06
  • 网络出版日期:  2022-06-06

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