摘要
本文提出了一个改进了的大麦黄花叶病毒提纯方法。病大麦叶片在高浓度(0.5M)磷酸钾缓冲液pH7.0(内含0.1%ME,0.01M EDTA)中匀浆,经1/4体积四氯化碳澄清后,病汁液用6%PEG,3%NaCl和1%TritonX-100混合物沉淀,高浓度(0.5M)同种缓冲液(含0.5M尿素,0.1%ME和0.01M EDTA)悬浮,接着进行20%蔗糖垫(含0.3%Triton X-100)超迷离心去除寄主细胞成分,10—40%蔗糖密度梯度离心进一步纯化。所获得的BaYMv提纯制剂A260/A280,A260/A240比值分别为1.20和1.04,纯病毒产量约为5.5—8.0mg/kg病叶。提纯病毒制剂在电镜下看不到有寄主杂质存在。
A purification procedure of barley yellow mosaic virus(BaYMV)was impro-ved:The infected barley leaves were homogenized with high molarity(0.5M)pota-ssium phosphate buffer pH7.0 containing 0.1% ME and 0.01M EDTA.After cla-rification with1/4 V carbon tetrachloride,the virus was concentrated by precip-itation with 6% PEG and 3%NaCl added 1% Triton x-100,and resuspended with the same buffer containing 0.5M urea,0.1% ME and 0.01M EDTA,followed by ul-tracentrifugation with a 20% suctose cushion added 0.3% Triton X-100,and futher purified by 10—40% sucrose density gradient centrifugation.The purified BaYMV thus obtained showed the following characteristics:A260/A280 ratio of 1.20,Max.260/Min.240 ratio of 1.04.The yield of purifed BaYMVwas about 5.5—8.0mg/kg infected leaves.Not any contaminats from host materials were observed in the pu-rified preparation under electron microscopy.
出处
《植物病理学报》
CAS
CSCD
北大核心
1989年第1期35-39,共5页
Acta Phytopathologica Sinica
