文献类型：期刊文献

中文题名：罗非鱼SRA4基因重组表达、亚细胞定位及组织分布

英文题名：Molecular Cloning,Subcellular Localization and Expression Characterization of SRA4 Gene in Nile Tilapia(Oreochromis niloticus)

作者：黄子威[1,2,3,4,5];汪志文[4,5];黎源[1,2,3,4,5];夏洪丽[4,5];简纪常[1,2,3];鲁义善[1,2,3,4,5]

机构：[1]广东海洋大学水产学院;[2]广东省水产经济动物病原生物学及流行病学重点实验室;[3]广东省水产经济动物病害控制重点实验室,广东湛江524088;[4]广东海洋大学深圳研究院;[5]广东省水生动物健康评估工程技术研究中心,广东深圳518000

年份：2021

卷号：41

期号：5

起止页码：19

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：CSTPCD、、北大核心、北大核心2020

基金：广东省现代农业产业体系项目(2019KJ141);深圳市科技计划项目(JCYJ20180306173023022);深圳市大鹏新区科技创新和产业发展专项资金项目(PT202101-23)。

语种：中文

中文关键词：尼罗罗非鱼;On-SRA4;重组表达;RT-qPCR;亚细胞定位

外文关键词：Oreochromis niloticus;On-SRA4;recombinant expression;RT-qPCR;subcellular localization

中文摘要：【目的】研究尼罗罗非鱼(Oreochromis niloticus)清道夫受体基因(On-SRA4)的重组表达、亚细胞定位及组织分布,为深入研究On-SRA4基因功能奠定基础。【方法】根据NCBI上公布的On-SRA4基因序列设计克隆引物,构建原核表达载体并诱导重组蛋白rOn-SRA4的表达。同时,采用生物信息学、亚细胞定位手段分析On-SRA4的结构特征,用实时定量PCR分析On-SRA4在头肾、胸腺、脾脏、肝、肠道、鳃、脑、肌肉、皮肤、血液的表达水平,用灭活无乳链球菌(Streptococcus agalactiae)、病毒类似物Poly I:C刺激健康罗非鱼,分析On-SRA4在头肾、胸腺、脾脏、肠道的表达水平。【结果与结论】成功构建原核表达载体pET28a-OnSRA4,诱导获得目的蛋白rOn-SRA4,rOn-SRA4分子质量约80 ku。亚细胞定位显示,rOn-SRA4定位于细胞核与细胞质。组织定量分析表明,On-SRA4在所有组织中均有表达,且在血液中表达量最高,在灭活无乳链球菌与Poly I:C刺激后,健康罗非鱼胸腺、肠道、脾脏和头肾中On-SRA4表达量均显著升高,表明On-SRA4参与了病原引起的免疫反应,是罗非鱼抵抗病原的相关基因。

外文摘要：【Objective】To study the restructuring expression,subcellular localization and tissue distribution of the scavenging receptor gene SRA4(On-SRA4)of Nile tilapia,Oreochromis niloticus,and provide information for further study on the function of On-SRA4.【Methods】Primers were designed according to gene sequence of On-SRA4 published on NCBI,and the prokaryotic expression vector was constructed to produce recombinant protein for rOn-SRA4.Also,bioinformatics analysis and subcellular localization were used to characteristize On-SRA4 by real-time quantitative PCR.Healthy tilapia were stimulated by inactivated Streptococcus agalactiae and virus analogs Poly I:C,and the expression levels of On-SRA4 in head kidney,thymus,spleen,intestine were analyzed.【Results and Conclusion】The prokaryotic expression vector pET28a-OnSRA4 was constructed,and a 80 ku rOn-SRA4 was induced.The results showed that On-SRA4 was localized in the nucleus and cytoplasm.Tissue quantitative PCR results showed that On-SRA4 was expressed in many tissues,and the highest expression level was found in the blood.After stimulation of healthy tilapia with inactivated S.agalactiae and Poly I:C,the expression level of On-SRA4 was significantly increased in the thymus,intestinal tract,spleen and head kidney.On-SRA4 is involved in the immune response caused by pathogens and is the gene related to resistance to pathogens.