从草鱼细胞分离到一种抗出血病病毒蛋白因子的研究  

STUDIES ON AN ANTI-HEMORRHAGIC VIRUS PROTEIN ISOLATED FROM CULTURE CELL OF GRASS CARP

作　　者：邵健忠[1] 项黎新[1] 李亚南[1] 毛树坚[1] 

机构地区：[1]杭州大学生物研究所,杭州310012

出　　处：《病毒学报》1993年第4期352-360,共9页Chinese Journal of Virology

基　　金：国家863计划生物技术领域资助

摘　　要：从人工诱变筛选的草鱼抗出血病病毒AHZC88细胞中检测到一种高效的抗病毒因子,经(NH_4)_2SO_4沉淀,PAGE洗脱和HPLC分离,得到纯化的抗病毒蛋白。该蛋白在出血病病毒的敏感细胞株ZC7901和CP80上表现出很强的抗病毒活性,平均滴度Log_4CPEI_(50)/0.1 ml在7.10±0.10(±S)以上。理化性质研究表明,该因子100000g离心不沉降,抗酸,不耐热,等电点为5.2。转译后分子量为29kD,经糖基化修饰成为38kD和36kD两种糖肽,未检出明显的磷酸化修饰,其沆病毒作用依赖于细胞内RNA和蛋白质的合成,并存在一定的细胞或病毒特异性。该因子在鱼体中有明显的抗病毒感染作用,能降低发病率,延缓发病时间。An anti-hemorrhagic virus protein factor was detected in a virus re-sistant cell line AHZCS8 of Grass Carp which was established by use of UV mutagenesis and selection. This protein factor was obtained by means of ( NH4)2SO4 sedimentation, Polyacrylamid gel electrophoresis and high performance liquid chromatography seperation. The average inhibitor titer(Log4 CPEI50/0.1ml) of the antiviral protein assayed on ZC7901 and CP80 cell lines sensitive to hemorrhagic virus of Grass Carp was higher than 7.10±0.10 (X±Sx) Iog4 units. Biochemical studies showed that the protein factor was not sedimented at 100,000g for 2 hours, exhibited sta bility to acid pH2 and unstability to heat 60℃. Its isoelectric point ( pI ) was about 5.2. The molecular weight of the antiviral protein precursor after being translated was 29, 000 daltons, through a post-translational glycosylation, it is converted into two distinct molecular forms of 38,000 and 36,000 daHons. The protein factor was not obviously phosphorylated. Inhibition of virai replication in celis is dependent on the process of cel-lular RNA transcription and protein synthesis. It was found that the protein factor was host or virus specific in its antiviral activity and was protective against infection by hemorrhagic virus when tested in the fin-gerlines of Grass Carp, as shown by the decline in mortality, and post-ponement of the time of death.

关 键 词：草鱼出血病 抗病毒蛋白 细胞 

分 类 号：S941.411[农业科学—水产养殖]