文章摘要
谷氨酰胺对GF-1细胞中C-Myc蛋白的表达与神经坏死病毒复制的关系研究
The study of the relationship between the expression of C-Myc protein and the replication of Nervous necrosis virus in GF-1 cells treated with
投稿时间:2019-09-17  修订日期:2019-10-25
DOI:
中文关键词: 斜带石斑鱼  C-Myc基因  GF-1细胞  神经坏死病毒  感染  谷氨酰胺
英文关键词: Epinephelus coioides  C-Myc gene  GF-1 cell  Nervous necrosis virus  Infection  Glutamine
基金项目:国家自然科学基金
作者单位E-mail
沈海洋 华南师范大学生命科学学院 953650617@qq.com 
卢志杰 华南师范大学生命科学学院  
秦真东 仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心  
赵丽娟 仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心  
李亚男 仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心  
刘春 仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心  
杨光 华南师范大学生命科学学院  
叶成凯 华南师范大学生命科学学院  
易敢峰 华南师范大学生命科学学院  
林蠡 仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心 linli@zhku.edu.cn 
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中文摘要:
      为探讨C-Myc表达、谷氨酰胺代谢和神经坏死病毒复制三者之间的关系,本研究首先克隆了斜带石斑鱼鳍条细胞(GF-1)中的C-Myc基因(GF-1-C-Myc),结果显示GF-1-C-Myc 基因cDNA全长814 bp,开放阅读框(ORF)有285 bp,编码95个氨基酸(aa),有亮氨酸拉链结构域与螺旋-环-螺旋(HLH)结构域。表达和纯化了GF-1-C-Myc 蛋白,并制备其多克隆抗体。采用实时定量PCR技术(qRT-PCR) 与免疫印迹法(WB)检测了GF-1-C-Myc 基因的表达和神经坏死病毒的复制。结果显示,缺乏谷氨酰胺会同时抑制GF-1-C-Myc基因的表达和NNV的复制,添加谷氨酰胺可同时促进GF-1-C-Myc的表达和NNV的复制;此外,NNV感染可上调GF-1-C-Myc 基因的表达,并显著消耗GF-1 细胞培养液中的谷氨酰胺。研究表明,GF-1-C-Myc基因可调控宿主谷氨酰胺代谢,从而有利于神经坏死病毒的复制。本结果为防控神经坏死病毒的感染提供了参考。
英文摘要:
      In order to investigate the relationships among the C-Myc expression, glutamine metabolism and replication of NNV. First of all, the C-Myc gene (GF-1-C-Myc) from Epinephelus coioides fin cell (grouper fin cells, GF-1) was cloned. The full length of GF-1-C-Myc gene cDNA was 814bp with 285bp ORF, encoding 95 amino acid (aa) with leucine zipper domain and helix-ring-helix (HLH) domain. GF-1-C-Myc protein was expressed and purified, and its polyclonal antibody was generated. The expressions of GF-1-C-Myc gene and the replication of NNV were monitored by real-time quantitative PCR (qRT-PCR) and immunoblotting (WB). The results showed that lack of glutamine could inhibit both the expression of GF-1-C-Myc gene and replication of NNV, while additional glutamine could promote both the expression of GF-1-C-Myc gene and replication of NNV. In addition, the expression of GF-1-C-Myc gene was up-regulated in GF-1 cells infected with NNV, and the glutamine in the medium was significantly consumed. Taken together, GF-1-C-Myc gene was involved in the regulation of glutamine metabolism in the cell, subsequently facilitated the replication of NNV. Our results will shed a new light on the prevention and control of NNV infection.
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