庄明鸽, 江世贵, 周发林, 黄建华, 杨其彬, 姜松, 杨丽诗. 斑节对虾CFSH基因的克隆及其多功能性探究[J]. 南方水产科学, 2020, 16(4): 90-99. DOI: 10.12131/20200011
引用本文: 庄明鸽, 江世贵, 周发林, 黄建华, 杨其彬, 姜松, 杨丽诗. 斑节对虾CFSH基因的克隆及其多功能性探究[J]. 南方水产科学, 2020, 16(4): 90-99. DOI: 10.12131/20200011
ZHUANG Mingge, JIANG Shigui, ZHOU Falin, HUANG Jianhua, YANG Qibin, JIANG Song, YANG Lishi. Molecular cloning and multifunction exploration of CFSH gene in Penaeus monodon[J]. South China Fisheries Science, 2020, 16(4): 90-99. DOI: 10.12131/20200011
Citation: ZHUANG Mingge, JIANG Shigui, ZHOU Falin, HUANG Jianhua, YANG Qibin, JIANG Song, YANG Lishi. Molecular cloning and multifunction exploration of CFSH gene in Penaeus monodon[J]. South China Fisheries Science, 2020, 16(4): 90-99. DOI: 10.12131/20200011

斑节对虾CFSH基因的克隆及其多功能性探究

Molecular cloning and multifunction exploration of CFSH gene in Penaeus monodon

  • 摘要: 该研究通过RACE (Rapid-amplification of cDNA ends) 法克隆并获得斑节对虾 (Penaeus monodon) 甲壳动物雌性激素 (Crustacean female sex hormone, CFSH) 基因PmCFSH的开放阅读框(ORF)及3'非编码区(UTR),预测编码214 氨基酸(aa)的蛋白,其含有信号肽和1个与免疫应答相关的白介素17E (IL-17E) 结构域。qRT-PCR结果显示,PmCFSH基因在各个组织中均有表达,其中在腹神经组织中的表达量最高;从受精卵时期到仔虾期的表达量呈逐渐上升趋势,其中在仔虾期表达量最高;在卵巢发育Ⅱ—Ⅳ期,随卵巢发育成熟表达量逐渐升高;革兰氏阴性菌和阳性菌均能上调PmCFSH基因的表达量,其中鳃组织PmCFSH在哈维氏弧菌 (Vibrio harveyi) 感染后第3小时和金黄色葡萄球菌 (Staphylococcus aureus) 刺激后第12小时达到表达高峰;而肝胰腺PmCFSH在哈维氏弧菌感染后第3小时和金黄色葡萄球菌刺激后第48小时达到表达高峰。研究表明,PmCFSH基因不仅参与幼体发育以及性腺成熟的调控,对细菌的应激也具有免疫应答响应,体现了激素的“多功能性”现象。

     

    Abstract: We cloned the ORF and 3'UTR of PmCFSH Crustacean female sex hormone (CFSH) gene of Penaeus monodon by RACE (Rapid-amplification of cDNA ends) method. It encodes a protein of 214 aa which contains a signal peptide and an interleukin 17E (IL-17E) domain relating to immune response. The qRT-PCR results showed that PmCFSH was expressed in various tissues, among which the highest expression level was found in the abdominal nerve tissue. The expression level gradually increased from fertilized egg stage to larval stage, and reached the maximum at larval stage. During the ovarian development Stage II to IV, it gradually increased as the ovary matured. Both Gram-negative and positive bacteria could increase the PmCFSH expression which was maximum in gill at 3rd hour after Vibrio harveyi infection and 12th hour after Staphylococcus aureus stimulation, while that in hepatopancreas reached the maximum at 3rd hour after V. harveyi infection and 48th hour after S. aureus stimulation. The study reveals that PmCFSH gene not only participates in the regulation of larval development and gonad maturity, but also has an immune response to bacterium, reflecting the "multifunction" of hormones.

     

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