中国水产科学Issue(5)：908-915,8.DOI:10.3724/SP.J.1118.2015.14448
草鱼天然抗性相关巨噬蛋白基因的表达特征与蛋白表达
Expression characteristics and protein expression of the natural resis-tance associated macrophage protein (Nramp) gene in grass carp (Ctenopharyngodon idellus)
摘要
Abstract
To explore the function of Nramp in the grass carp (Ctenopharyngodon idellus) immune system, gene expression was analyzed in different developmental stages and tissues. Four tissues of grass carp at different times after challenge with the pathogenic bacterium Aeromonas hydrophila were analyzed by real-time quantitative RT-PCR. The 642 bp Open Reading Frame (ORF) region of grass carp Nramp was expressed using prokaryotic expression technology. The fusion protein had a relative molecular mass of 40 kD. Real-time quantitative RT-PCR analysis indicated that the Nramp mRNA level in embryos gradually increased during embryogenesis from cleav-age to fry stages after fertilization, with the highest level in the fry stage. In adult fish, the Nramp mRNA level in different tissues significantly differed; Nramp transcripts were found to be highly abundant in liver; moderately abundant in the head kidney, spleen, and intestine;and least abundant in the eye and bladder. After challenge with A. hydrophil, with uninjected groups as the control, the Nramp mRNA level in the head kidney, liver, spleen, and intestine significantly increased (P<0.05). There was significant up-regulation in head kidney at 12 h post-treatment, with a maximum level at 48 h post-infection;significant down-regulation at 72 h and 96 h;and the expression regressed to the initial level by 7 d. Nramp expression in the liver was significantly increased at 12 h post-treatment;followed by a decrease at 24 h post-treatment;another increase at 48 h, with a maximum level at 72 h post-infection;and the high expression level was maintained at 96 h and 7 d post-treatment. There was sig-nificant up-regulation in the intestine at 12 h post-treatment, with a maximum level at 48 h post-treatment; sig-nificant down-regulation at 72 h and 96 h; and the expression regressed to the initial level by 7 d. In the spleen, Nramp mRNA level increased at 4 h post-treatment, with a maximum level at 12 h; then, there was significant down-regulation at 24 h and 48 h;and the expression regressed to the initial level by 7 d. Bacterial infection can increase Nramp expression in immune-related organs, which may indicate that Nramp plays an important role in the innate immune response of grass carp.关键词
草鱼/Nramp基因/表达分析/原核表达/蛋白纯化Key words
Ctenopharyngodon idellus/Nramp gene/expression and analysis/prokaryotic expression/protein purification分类
农业科技引用本文复制引用
杨康,王艺雅,刘铁柱,李同明,季相山,王慧..草鱼天然抗性相关巨噬蛋白基因的表达特征与蛋白表达[J].中国水产科学,2015,(5):908-915,8.基金项目
国家科技支撑计划项目(2012BAD26B00) （2012BAD26B00）
山东省优秀中青年科学家科研奖励基金项目(BS2013NY002) （BS2013NY002）
山东省现代农业产业技术体系项目(SDAIT-15-011) （SDAIT-15-011）
