草莓伪温和黄边病毒提纯技术的研究

Study on the Purification Techniques for Strawberry Pseudo-Mild Yellow Edge Virus

纯化的草莓伪温和黄边病毒(SPMYEV),小叶嫁接繁殖于指示植物草莓UC—4上,然后采样榨汁,结合聚乙二醇(PEG)沉淀,蔗糖垫底差速离心及连续性蔗糖密度梯度离心,能成功地将草莓伪温和黄边病毒提纯出来。经电镜观察为丝杆状病毒粒体,长×宽为650nm×12—13nm。提纯的病毒制剂,经紫外分光光度计测定,呈典型的核蛋白吸收曲线,其最大值位于263nm,最小值位于243nm。A260/A280=1.24,用冰醋酸分解病毒,提取衣壳蛋白,经SDS-聚丙烯酰胺凝胶电泳测定其分子量约37000道尔顿。

The isolated strawberry pseudo-mild yellow edge virus(SPMYEV)was maintained in the strawberry-Fragaria vesca L.var. UC-4, and was successfully extracted and purified through homogenization by polyethyleneglycol (PEG) precipitation, differential centrifugation in the sucrose cushion and continuous suerose density gradient centrifugation. The purified preparations cotained flexous rod-shaped virus particles mainly 650nm in length and 12—13nm in width. The virus showed a typitical UV spectrum of nueleoprotein with max. and min. absorption at 263nm and 243nm respectively. The A260/A280=1.24. The coat protein of SPMYEV was obtained by using cold acetic acid, and had one band with the mol.wt.37000 daltons(average of four determinations), it was through the SDS-polyacrylamide gel electrophoresis plate.