论文部分内容阅读
在甜菜坏死黄脉病毒分离提纯的研究中,获得了一个较好的提纯程序。用刚显症的新鲜番杏病叶,加PH8.4、0.2m的硼酸缓冲液(内含0.5％巯基乙醇,0.1m脲素、0.005mEDTA),捣碎过滤,6％PEG沉淀后,通过SePharose2B柱层析染纯化病毒。病毒经6％PEG再次沉淀浓缩后,调节病毒悬液PH至酸性使病毒易悬浮,并通过病毒等电点沉淀法进一步纯化病毒。纯化病毒制剂呈典型的病毒核蛋白紫外吸收曲线,最大吸收值在270毫微米,最小吸收值在250毫微米。该病毒分离物OD_(260)/OD_(280)为1.123左右,核酸含量约4.5％,病毒等电点PH4.8～4.9。病毒的核酸电泳出现4条带,它们的分子量约2.25×10~6、1.8×10~6、1.05×10~6、0.75×10~6道尔顿。病毒粒体含一种外壳蛋白亚基,分子量约2.05×10~6道尔顿,由约16种199个氨基酸组成。病毒制剂在分析超速离心时出现了4个沉降界面,它们各自的沉降系数分别为200.8S,165S,125.8S和100S。
In the study of separation and purification of beet necrotic vein virus, a better purification procedure was obtained. With fresh symptoms of fresh apricot disease leaves, add PH8.4, 0.2m borate buffer (containing 0.5% mercaptoethanol, 0.1m urea, 0.005mEDTA), mashed and filtered, 6% PEG precipitation, by SePharose2B column chromatography to purify the virus. The virus was again precipitated and concentrated by 6% PEG, and the pH of the virus suspension was adjusted to be acidic to make the virus easy to suspend. The virus was further purified by isoelectric point precipitation of virus. The purified virus preparation showed a typical ultraviolet absorption curve of the nucleocapsid protein with a maximum absorption value of 270 nm and a minimum absorption value of 250 nm. The OD260 / OD280 of the virus isolate was about 1.123, the nucleic acid content was about 4.5%, and the isoelectric point of the virus was 4.8-4.9. The virus nucleic acid electrophoresis appeared 4 bands, their molecular weight of about 2.25 × 10 ~ 6, 1.8 × 10 ~ 6, 1.05 × 10 ~ 6, 0.75 × 10 ~ 6 Dalton. The virus genome contains a coat protein subunit with a molecular weight of about 2.05 × 10 ~ 6 Daltons and consists of about 16 199 amino acids. Four subsidence interfaces of the virus preparation were observed during the analysis of ultracentrifugation, and their respective sedimentation coefficients were 200.8S, 165S, 125.8S and 100S, respectively.