论文部分内容阅读
通过对番茄环斑病毒繁殖寄主、抽提缓冲液、澄清剂、浓缩方法和悬浮缓冲液等比较试验,初步筛选出得率高,并能保持病毒完整的提纯程序,即:用番杏为繁殖寄主,用0.05M 磷酸钾缓冲液(pH 7.5,内含0.02M 巯基乙醇)抽提,加1%Tritonx-100澄清,经8%聚乙二醇沉淀,接着差速离心(28000转/分,4小时)浓缩病毒,最后用0.01 MPB(pH 7.5内含0.002MEDTA)悬浮,低速离心,上清液为部分提纯的病毒制剂。此提纯制剂具有侵染性;病毒粒体完整,为等轴对称,直径约28μm;且具有典型的核蛋白吸收曲线。
Through the comparative test of tomato ringspot virus breeding host, extraction buffer, clarifying agent, concentration method and suspension buffer, we can select the high purification rate and maintain the integrity of the virus, namely: Host, extracted with 0.05 M potassium phosphate buffer pH 7.5 containing 0.02 M mercaptoethanol, clarified with 1% Tritonx-100, precipitated with 8% polyethylene glycol, followed by differential centrifugation (28,000 rpm, 4 hours) to concentrate the virus, finally suspended in 0.01 MPB (containing 0.002MEDTA in pH 7.5), centrifuged at low speed, and the supernatant was partially purified. The purified preparation is infectious; the virus is intact, equiaxed and has a diameter of about 28 μm; and has a typical nuclear protein absorption curve.