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目的 :探索一条既能获得dsRNA病毒全基因组 ,又能同时获得该病毒完整结构多肽的方法。方法 :将分别感染了蓝舌病毒 (BTV)和人轮状病毒 (HRV)的培养细胞分别破碎以释出病毒后 ,结合应用差速离心和蔗糖梯度离心技术 ,获得了这两种病毒的纯化粒子 ;再用 1%的SDS裂解病毒 ,0 .2 5mol/L的KCl沉淀其结构蛋白 ,而病毒基因组悬于上清。结果 :所沉淀的蛋白用PBS(pH 7.8)适当稀释后 ,即可用作抗原 ;上清用酚∶仿抽提 1次后 ,即可用于核酸分子生物学研究。结论 :此方法为一套既能有效同步分离dsRNA病毒核酸和结构多肽等活性生物大分子 ,又能节约实验材料和时间的有效方法。
OBJECTIVE: To explore a method to obtain both the whole genome of dsRNA virus and the complete structural polypeptide of the virus simultaneously. Methods: The cultured cells were infected with BTV and HRV, respectively. After the virus was released, the two viruses were purified by differential centrifugation and sucrose gradient centrifugation The particles were then lysed with 1% SDS and the structural protein was precipitated with 0.25 mol / L KCl while the viral genome was suspended in the supernatant. Results: The precipitated protein was diluted with PBS (pH 7.8) and used as antigen. After the supernatant was extracted with phenol for 1 time, it could be used for nucleic acid molecular biology research. Conclusion: This method is an effective method for simultaneous separation of active biological macromolecules such as dsRNA virus nucleic acid and structural polypeptide, as well as the saving of experimental materials and time.