水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2014年
3期
430-437
,共8页
张凤%李周全%闫利明%罗绍祥%钟利桥%张晓华%袁丽%方勤%戴和平
張鳳%李週全%閆利明%囉紹祥%鐘利橋%張曉華%袁麗%方勤%戴和平
장봉%리주전%염리명%라소상%종리교%장효화%원려%방근%대화평
草鱼呼肠孤病毒%VP5%VP7%噬菌体展示%单链抗体
草魚呼腸孤病毒%VP5%VP7%噬菌體展示%單鏈抗體
초어호장고병독%VP5%VP7%서균체전시%단련항체
Reovirus of grass carp (GCRV)%VP5%VP7%Phage display%scFv antibody
草鱼呼肠孤病毒(GCRV)是引起我国大面积草鱼幼鱼出血病暴发的主要病原,其外衣壳蛋白 VP5和VP7在病毒入侵宿主细胞过程中起着至关重要的作用。研究以原核表达的VP7、全长VP5、VP5的N端片段及C端片段为靶蛋白,利用已构建的噬菌体展示单链抗体文库进行淘选。经过3轮淘选后,共获得7个针对VP7、VP5、VP5N和VP5C的单链抗体。经过验证,识别原核表达的VP7的两个单链抗体能够成功识别天然GCRV病毒。此结果对于进一步研究GCRV与宿主细胞的相互作用机理奠定了基础。
草魚呼腸孤病毒(GCRV)是引起我國大麵積草魚幼魚齣血病暴髮的主要病原,其外衣殼蛋白 VP5和VP7在病毒入侵宿主細胞過程中起著至關重要的作用。研究以原覈錶達的VP7、全長VP5、VP5的N耑片段及C耑片段為靶蛋白,利用已構建的噬菌體展示單鏈抗體文庫進行淘選。經過3輪淘選後,共穫得7箇針對VP7、VP5、VP5N和VP5C的單鏈抗體。經過驗證,識彆原覈錶達的VP7的兩箇單鏈抗體能夠成功識彆天然GCRV病毒。此結果對于進一步研究GCRV與宿主細胞的相互作用機理奠定瞭基礎。
초어호장고병독(GCRV)시인기아국대면적초어유어출혈병폭발적주요병원,기외의각단백 VP5화VP7재병독입침숙주세포과정중기착지관중요적작용。연구이원핵표체적VP7、전장VP5、VP5적N단편단급C단편단위파단백,이용이구건적서균체전시단련항체문고진행도선。경과3륜도선후,공획득7개침대VP7、VP5、VP5N화VP5C적단련항체。경과험증,식별원핵표체적VP7적량개단련항체능구성공식별천연GCRV병독。차결과대우진일보연구GCRV여숙주세포적상호작용궤리전정료기출。
Grass carp reovirus (GCRV) is the main cause of severe outbreaks of hemorrhagic disease in fingerling and yearling grass carp. The virus outer capsid proteins, VP5 and VP7, have been shown to play critical roles in the invasion of host cells. In our study, prokaryotic-expressed VP7, VP5, and the N and C terminals of VP5 protein were used as targets to select the desired antibodies from phage-displayed scFv library. After 3 rounds of panning, we selected 7 an-tibodies against the prokaryotic-expressed antigens-- VP7, VP5, VP5N and VP5C. Furthermore, we found that 2 anti-bodies against VP7 out of the total 7 were able to recognize native GCRV particles. Our study should provide an insight into the interactions between GCRV and its host cells.