文献类型：期刊文献

中文题名：尼罗罗非鱼跨膜Bax抑制剂母体6基因Tmbim 6的克隆、表达及功能鉴定

英文题名：Molecular cloning,expression and functions of Tmbim 6 gene in Nile tilapia (Oreochromis niloticus)

作者：巫祎琴[1];陈福暖[1];张志强[1];江栢坚[1];简纪常[1];黄瑜[1]

机构：[1]广东海洋大学水产学院,广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088

年份：2023

卷号：38

期号：2

起止页码：275

中文期刊名：大连海洋大学学报

外文期刊名：Journal of Dalian Ocean University

收录：CSTPCD、、CSCD2023_2024、北大核心、CSCD、北大核心2020

基金：国家自然科学基金(32002426,32073006);湛江市非资助科技攻关计划项目(2020B01169)。

语种：中文

中文关键词：尼罗罗非鱼;Tmbim 6;亚细胞定位;NF-κB;基因表达

外文关键词：Oreochromis niloticus;Tmbim 6;subcellular localization analysis;NF-κB;expression analysis

中文摘要：为了解跨膜Bax抑制剂母体6(transmembrane Bax inhibitor motif containing 6,TMBIM 6)在尼罗罗非鱼(Oreochromis niloticus)病原体感染中的作用,利用聚合酶链式反应(PCR)对Tmbim 6基因进行克隆与鉴定,并采用实时荧光定量PCR(qPCR)分析Tmbim 6在健康尼罗罗非鱼(体质量为80~100 g)的组织分布模式及无乳链球菌(Streptococcus agalactiae)、Poly I:C模拟病毒刺激后的表达变化。结果表明:尼罗罗非鱼Tmbim 6开放阅读框为714 bp,可编码237个氨基酸,包含6个跨膜结构域和1个C-末端基序;亚细胞定位显示,尼罗罗非鱼TMBIM 6定位于细胞质,双荧光素酶报告基因系统检测发现,Tmbim 6在HEK-293T细胞中过表达后极显著抑制NF-κB通路(P<0.01);qPCR分析显示,Tmbim 6基因在所检测的组织中广泛分布,在肝脏和肌肉中的表达量最高;经无乳链球菌感染和Poly I:C刺激后,Tmbim 6在肝脏、脾脏、头肾、脑和肠道中的表达水平均显著上调(P<0.05)。研究表明,尼罗罗非鱼TMBIM 6参与了无乳链球菌感染和Poly I:C刺激后的细胞应激过程及免疫反应。

外文摘要：To understand the role of transmembrane Bax inhibitor motif containing 6(Tmbim 6 gene) in pathogen infection of Nile tilapia(Oreochromis niloticus),the Tmbim 6 gene was cloned,identified and expressed in healthy Nile tilapia(body weight 80-100 g) induced by Poly I:C and Streptococcus agalactiae by quantitative real-time PCR.Results showed that the Tmbim 6 gene had 714 bp ORF encoding a protein of 237 aa,and containing six transmembrane regions and C-terminus motif.Subcellular localization analysis indicated that the Tmbim 6 gene was located in the cytoplasm and Dual-luciferase reporter assays showed that over-expression of Tmbim 6 in HEK-293T cell suppressed the NF-κB reporter gene(P0.01).qPCR analysis revealed that Tmbim 6 was widely distributed in different tissues of healthy tilapia,with the maximal expression level in the liver and the minimal expression level in the head kidney and spleen.Expressions of Tmbim 6 gene were found to be significantly up-regulated in liver,spleen,intestine,head kidney and brain in response to Poly I:C and Streptococcus agalactiae stimulations(P0.05).In conclusion,the present findings revealed that Nile tilapia Tmbim 6 gene was involved in the cell stress response and immune process in response to pathogen infection.