草鱼TRIM32基因克隆表达及功能初步研究
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- 英文论文题名：The preliminary study on cloning, expression profiles and function of grass carp(Ctenopharyngodon idellus)TRIM32
- 论文作者：贾路路 ; 周勇 ; 王方 ; 彭俊杰 ; 王业大 ; 曾令兵 ; 刘学芹
- 英文论文作者：JIA Lulu ; ZHOU Yong ; WANG Fang ; PENG Junjie ; WANG Yeda ; ZENG Lingbing ; LIU Xueqin ; College of Fisheries ; Huazhong Agricultural University ; Chinese Academy of Fishery Sciences ; Yangtze River Fisheries Research Institute
- 年：2016
- 作者机构：华中农业大学水产学院;中国水产科学研究院长江水产研究所;
- 论文关键词：TRIM32 ; 克隆 ; 真核表达 ; NF-κB
- 英文论文关键词：TRIM32 ; cloning ; eukaryotic expression ; NF-κB
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：577k
- 原文格式：D
- 会议级别：全国
摘要
TRIM(tripartite-motif protein)家族是机体先天性免疫的重要成员,参与细胞增殖分化、细胞凋亡、肿瘤抑制、抗病毒等过程。本实验利用PCR方法克隆了草鱼TRIM32基因的编码区全长,共1980bp,编码660个氨基酸。草鱼TRIM32具有TRIM家族典型的3个结构域,与斑马鱼TRIM32的核苷酸序列同源性较高,遗传进化分析也聚为一支。间接免疫荧光、Western-blotting检测结果显示草鱼TRIM32在EPC细胞中成功表达,主要以点状分布在细胞质中,细胞核中表达量较少;组织分布分析表明TRIM32在被检测的11个组织中均有表达,其中在头肾组织中的表达量明显高于其他组织;不同胚胎发育时期的表达分析进一步表明,TRIM32在受精卵时期、卵裂期和囊胚期的表达量显著高于其他时期,随后表达量下降,直到出膜后表达量再次显著性升高。此外,双荧光素酶检测系统显示TRIM32能够激活NF-κB信号通路,诱导下游的炎症反应。
The TRIM family is an important member in natural immune system, which involved in cell proliferation, differentiation, apoptosis, tumor suppression, antiviral activity and other processes. The full-length coding sequence region of TRIM32 from grass carp was cloned using PCR method. It was 1980 bp in length encoding 660 amino acid.The TRIM32 protein has three typical structural featuresof the TRIM family. Sequence analysis indicated that TRIM32 showed the highest identity with zebrafish and the phylogenetic trees converge the same branch.Immunofluorescence and Western-blotting assay showed that TRIM32 successfully expressed in EPC cells and localized in cytoplasmic speckles; tissue distribution analysis showed that TRIM32 was widely expressed in various tissues, especially in the renal tissue; different stages of embryonic development shown that in zygote period, cleavage and blastula stage TRIM32 expression were significantly higher than other times, and then decreased until hatching increased significantly again.
The TRIM family is an important member in natural immune system, which involved in cell proliferation, differentiation, apoptosis, tumor suppression, antiviral activity and other processes. The full-length coding sequence region of TRIM32 from grass carp was cloned using PCR method. It was 1980 bp in length encoding 660 amino acid.The TRIM32 protein has three typical structural featuresof the TRIM family. Sequence analysis indicated that TRIM32 showed the highest identity with zebrafish and the phylogenetic trees converge the same branch.Immunofluorescence and Western-blotting assay showed that TRIM32 successfully expressed in EPC cells and localized in cytoplasmic speckles; tissue distribution analysis showed that TRIM32 was widely expressed in various tissues, especially in the renal tissue; different stages of embryonic development shown that in zygote period, cleavage and blastula stage TRIM32 expression were significantly higher than other times, and then decreased until hatching increased significantly again.
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