摘要
用来自于白车根草 (Trifolium repens)上的一个苜蓿花叶病毒分离物 AMV- SY为材料 ,比较了 3种以差速离心为主结合 PEG沉淀和超速离心提纯病毒的方法 ,对提纯病毒进行紫外吸收测定、电镜检查和SDS-聚丙烯酰胺凝胶电泳检测的结果显示 :以交替使用含有 0 .1mol/LEDTA和 0 .1mol/L Mg SO4 的磷酸缓冲液作为病毒悬浮介质的提纯程序最为理想。该方法提取苜蓿花叶病毒的得率为 4 7.6 mg/10 0 g昆诺藜鲜病叶 ,该病毒分离物的外壳蛋白分子量为 2 9k D。该方法的病毒得率较高、杂蛋白较少、病毒粒子完整 ,是比较理想的提纯方法。
Three purification methods,based on differential centrifugation,precipitation by polyethylene glycol (PEG)and ultra speed centrifugation,were compared for purification of an alfalfa mosaic virus (AMV SY)previously isolated from Trifolium repens.The Purified virus was observed under electron microscope, measured,by ultra violet absorpton analysis and protein determination with SDS polyacrylamide gel electrophoresis.Our results showed that a method by alternative use of sodium phosphate buffers containing 0.1mol/L EDTA and 0.1mol/L MgSO 4 achieved the best purification with less miscellaneous protein contamination,integrate virus particles and relatively high yield, which was of 47.6mg virion per hundred grams of fresh leaves of Chanopodium quinoa inoculated with AMV SY.The coat protein of AMY SY was tested for about 29 kilo Dalton.
出处
《微生物学通报》
CAS
CSCD
北大核心
2001年第1期68-72,共5页
Microbiology China
基金
国家自然科学基金资助项目!(No.39880 0 2 8)&&
