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国家自然科学基金(31172434)

作品数:5 被引量:13H指数:3
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High-resolution 3D Structures Reveal the Biological Functions of Reoviruses被引量:3
2013年
Viruses in the family Reoviridae are non-enveloped particles comprising a segmented double-stranded RNA genome surrounded by a two-layered or multi-layered icosahedral protein capsid.These viruses are classified into two sub-families based on their particle structural organization.Recent studies have focused on high-resolution three-dimensional structures of reovirus particles by using cryo-electron microscopy (cryo-EM) to approach the resolutions seen in X-ray crystallographic structures.The results of cryo-EM image reconstructions allow tracing of most of the protein side chains,and thus permit integration of structural and functional information into a coherent mechanism for reovirus assembly and entry.
Xiaoming LiQin Fang
关键词:呼肠孤病毒X射线晶体结构病毒颗粒冷冻电镜
斑马鱼ftr56基因克隆表达及功能研究被引量:1
2020年
为进一步了解硬骨鱼类特有的finTRIM(Fish novel tripartite motif,ftr)在斑马鱼(Danio rerio)抗病毒天然免疫中的作用,研究克隆了斑马鱼ftr56基因并分析了其对鲤春病毒血症病毒(Spring viremia of carp virus,SVCV)增殖的抑制作用。根据NCBI中斑马鱼ftr56序列设计引物,采用PCR方法,扩增ftr56 CDS区,连接至真核表达载体pcDNA4.0-His,构建真核表达质粒pcDNA4.0-ftr56-His,进行生物信息学分析。采用实时荧光定量PCR(qRT-PCR)技术检测SVCV感染斑马鱼胚胎成纤维细胞(ZF4)后ftr56 mRNA的变化。系统进化树分析显示,斑马鱼FTR56单独聚为一支。氨基酸多序列比对结果显示,其与黑猩猩、牛、鼠的TRIM56相似度为22%—23%。FTR56二级结构具有1个RING指结构域,1个B-box结构域,1个卷曲螺旋结构域和1个B30.0结构域。qRT-PCR检测结果显示,ftr56在SVCV感染后24h表达量显著上升。在过表达ftr56后,SVCV的G蛋白mRNA水平和蛋白水平在12h和24h相比对照组明显减少,培养基上清中SVCV滴度也明显降低,表明FTR56抑制SVCV的增殖。实验为进一步揭示finTRIM在鱼类病毒性疾病中的免疫调节机制提供参考。
邝鸣刘晥蒙姚健霍诗天刘学芹
关键词:克隆表达抗病毒斑马鱼
Antigenic analysis of grass carp reovirus using single-chain variable fragment antibody against IgM from Ctenopharyngodon idella被引量:6
2013年
Grass carp (Ctenopharyngodon idella) is an important species of freshwater aquaculture fish in China. However, grass carp reovirus (GCRV) can cause fatal hemorrhagic disease in yearling populations. Until now, a strategy to define the antigenic capacity of the virus's structural proteins for preparing an effective vaccine has not been available. In this study, some single-chain variable fragment antibodies (scFv), which could specifically recognize grass carp IgM, were selected from a constructed mouse na ve antibody phage display cDNA library. The identified scFv C1B3 clone was shown to possess relatively higher specific binding activity to grass carp IgM. Furthermore, ELISA analysis indicated that the IgM level in serum from virus-infected grass carp was more than two times higher than that of the control group at 5-7 days post infection. Moreover, Western blot analysis demonstrated that the outer capsid protein VP7 has a specific immuno-binding-reaction with the serum IgM from virus-infected grass carp. Our results suggest that VP7 can induce a stronger immune response in grass carp than the other GCRV structural proteins, which implies that VP7 protein could be used as a preferred immunogen for vaccine design.
CHEN CongLinSUN XiaoYunLIAO LanJieLUO ShaoXiangLI ZhouQuanZHANG XiaoHuaWANG YaPingGUO QionLinFANG QinDAI HePing
关键词:草鱼呼肠孤病毒抗原性分析可变区VP7基因CDNA文库
N-Terminal Myristoylated VP5 is Required for Penetrating Cell Membrane and Promoting Infectivity in Aquareoviruses被引量:1
2018年
Dear Editor,Myristoylation is a naturally occurring post-translational modification for targeting cytoplasmic proteins to intracellular membranes.Unlike enveloped animal viruses,which enter host cells by membrane fusion,nonenveloped animal viruses must disrupt the cell membrane to initiate infection.Some animal viruses and several nonenveloped viruses
Qingxiu ChenHong GuoFuxian ZhangQin Fang
关键词:细胞膜传染性动物病毒
利用抗草鱼IgM的单链抗体分析草鱼呼肠孤病毒的免疫原性被引量:3
2012年
草鱼是中国重要的淡水养殖鱼种之一,草鱼呼肠孤病毒(GCRV)可引起一龄草鱼出现致命性的出血性疾病,严重影响草鱼养殖业的健康发展.目前,在草鱼中尚未建立一种有效的检测该病毒结构蛋白抗原性质的方法.本研究采用已构建的小鼠天然噬菌体展示抗体文库淘选得到可以特异性识别草鱼IgM的单链抗体,并在已鉴定的单链抗体阳性克隆中,筛选出对草鱼IgM有较好的特异性结合活性的单链抗体C1B3.ELISA分析表明,在感染草鱼出血病病毒5~7天后,感染组比对照组血清中IgM的含量增加了2倍多.进一步的Western blot结果显示,已感染病毒草鱼血清中的IgM能特异地识别草鱼的外壳蛋白VP7.结果表明,VP7相对于病毒的其他结构蛋白而言在草鱼中能产生更强的免疫反应,提示VP7应是用于疫苗设计的首选抗原.
陈丛琳孙小云廖兰杰罗绍祥李周全张晓华汪亚平郭琼林方勤戴和平
关键词:草鱼草鱼呼肠孤病毒免疫原性
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