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苜蓿花叶病毒提纯方法的改进*
引用本文:陈集双, 冯明光,.苜蓿花叶病毒提纯方法的改进*[J].微生物学通报,2001,28(1):68-72.
作者姓名:陈集双  冯明光  
作者单位:浙江大学生命科学学院
基金项目:国家自然科学基金资助项目(No.39880028)浙江省教委重点课题
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 MgSO4的磷酸缓冲液作为病毒悬浮介质的提纯程度最为理想,该方法提取苜蓿花叶病毒的得率为47.6mg/100g昆诺藜鲜病叶,该病毒分离物的外壳蛋白分子量为29kD。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。

关 键 词:苜蓿花叶病毒  白车根草  提纯方法  外壳蛋白  植物病毒
文章编号:0253-2654(2001)01-068-05
修稿时间:1999年10月27

MODIFIED TECHNIQUES FOR PURIFICATION OF ALFALFA MOSAIC VIRUS
CHEN Ji-shuang,FENG Ming-Guang.MODIFIED TECHNIQUES FOR PURIFICATION OF ALFALFA MOSAIC VIRUS[J].Microbiology,2001,28(1):68-72.
Authors:CHEN Ji-shuang  FENG Ming-Guang
Abstract: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 (AMVSY)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 MgSO4 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.
Keywords:Alfalfa mosaic virus  Trifolium repens  Purification method  Coat protein  
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