论文部分内容阅读
大白菜芜菁花叶病毒(TuMV)沈阳分离物,是本研究组分离和纯化的毒原。在无虫温室中以人工接种于大白菜(小白口)叶片,适期收获,称重,冰冻过夜。次日加适量0.02M磷酸缓冲液,pH7.2匀浆。并进行差速离心;低速离心3000rpm,30分钟去寄主细胞器,取上清液搅拌加4％聚乙二醇(PEG—MW6000)及3％NaCl(W/V),搅拌沉淀病毒粒体,进行一系列低速及超速离心。超速离心后的沉淀物用0.01M磷酸缓冲盐液(含0.015M尿素)悬浮,得到纯化的TuMV制品。经电子显微镜检查,提纯物中含有大量单个非集聚的TuMV粒体。粒体平均长度为720nm,结构保持良好,粒体断裂现象很少。提纯物经指示植物接种测定,具侵染性。这一简化提纯法可以免去蔗糖密度梯度离心等过程,取得较为相似的效果。
Chinese cabbage turnip mosaic virus (TuMV) Shenyang isolate, is the virulent source of isolation and purification in our group. In an insect-free greenhouse inoculated on Chinese cabbage (small white mouth) leaves, appropriate harvest, weighing, frozen overnight. The next day add appropriate amount of 0.02M phosphate buffer, pH7.2 homogenate. And centrifuged at low speed for 3000 rpm for 30 minutes to the host cell organelle. The supernatant was agitated with 4% polyethylene glycol (PEG-MW6000) and 3% NaCl (W / V) A series of low-speed and ultracentrifugation. The pellet after ultracentrifugation was suspended with 0.01 M phosphate buffered saline (containing 0.015 M urea) to obtain a purified TuMV preparation. Electron microscopy revealed that the purification contained a large number of individual non-aggregated TuMV granules. The average length of the mitochondria is 720 nm, the structure is well maintained, and the phenomenon of mitochondrial fragmentation is rare. Purified by the indicated plant inoculation assay, with infectivity. This simplified purification method can remove sucrose density gradient centrifugation and other processes to achieve a more similar effect.