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, 2022, 18(6): 52-59. 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, 2022, 18(6): 52-59. 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 from Macrobrachium rosenbergii[J]. South China Fisheries Science, 2022, 18(6): 52-59. 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 from Macrobrachium rosenbergii[J]. South China Fisheries Science, 2022, 18(6): 52-59. DOI: 10.12131/20220067

罗氏沼虾nanos1基因克隆、表达及亚细胞定位

Molecular cloning, expression profile and subcellular localization of nanos1 gene from 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序列全长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) 受精时期表达量最高,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。

     

    Abstract: To investigate the function of the nanos1 gene in the reproductive development of Macrobrachium rosenbergii, we cloned its nanos1 gene. The cDNA sequence was 2 811 bp, encoding 243 amino acids. Phylogenetic analysis shows that it had the highest homology with Eriocheir sinensis, higher homology with nanos1 in fish but lower homology with mammals. We detected the expression level of the gene at different embryonic developmental stages and in larvae by real-time fluorescence quantitative PCR. The results show that the expression level of nanso1 mRNA was the highest at unfertilized stage, significantly higher than that at fertilized stage and the other embryonic developmental stages (P<0.05). During the embryonic development, the expression was the highest at fertilization stage, significantly higher than that at cleavage stage and very significantly higher than that at late embryonic development (P<0.01). The expression level of this gene at cleavage stage was significantly higher than that from blastocyst stage to larval stage. The expression level from blastocyst stage to larval stage was low and there was no difference.In situ hybridization shows that nanos1 mRNA was expressed in the cytoplasm of oocytes and primary oocytes (Oc1, Oc2, Oc3 and Oc4). The results show that nanos1 is closely related to the development of female germ cells of M. rosenbergii.

     

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