水稻黄矮病毒结构蛋白的分离及其小鼠腹水抗体的制备

Studies on separation of structural proteins and production of ascite antibodies to the proteins of rice yellow stunt virus.

从人工接种感染黄矮病毒(Rice Yellow Stunt Virus,RYSV)的水稻茎杆中提取得到了部分纯化的RYSV制品,病毒制品经Tricine-SDS-PAGE得到分子量分别为84 kD,60 kD,31 kD和29.5 kD的4条主要蛋白带和分子量为170kD和43kD的两条次要蛋白带。根据弹状病毒结构蛋白命名法,按分子量大小依次命名为L,G,N,NS,M_1和M_2蛋白。从电泳凝胶上切下G和N蛋白带,制成抗原免疫BALB/C小鼠;制得G和N蛋白小鼠腹水抗体。经Dot-blot试验测得N蛋白抗体效价为1:1000,G蛋白抗体效价为1:400。

Rice yellow stunt virus (RYSV)is a member of plant rhabdo-viruses being composed of some structural proteins. The RYSV preparations from stalks of diseased plant by artificial inoculation were purified,and heat-treated in the presence of 1% SDS and 2% mercaptoethanol. Proteins were separated by Tricine-SDS-PAGE. After stained with coomassie brilliant blue,the preparations showed 4 major bands with the sizes of 84 kD,60 kD,31 kD and 29. 5kD and 2 minor bands with 170 kD and 43 kD. According to the nomenclature of structural proteins of rhabdoviruses they were mamed as L,G,N,NS,M, and M2 protein in the order of decreasing sizes. The antigens of G and N were prepared. The BALB/C mice were immunized by three subcutaneous injections with emulsified antigens and one boost intraperitoneal injection with soluble antigens .respectively. Later S180 cells were injected into abdominal cavities of mice and then ascite antibodies were extracted. The titers of G and N ascite in Dot-blot tests were 1 : 400 and 1 : 1 000,respectively.