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目的对人呼吸道合胞病毒(HRSV)的裂解条件进行初步探索,为研制HRSV裂解纯化疫苗奠定基础。方法用不同的裂解剂,以不同浓度和温度对HRSV裂解不同时间,以HRSV的F及G蛋白为目标蛋白,对得到的蛋白进行还原蛋白电泳分析及Western blot鉴定,电镜观察验证裂解效果。结果1%浓度的TritonX-100在4℃作用90min,对HRSV的裂解效果优于其他裂解剂。电镜观察可见病毒样品呈云絮状裂解;Western blot分析表明,特异性条带为HRSV的F蛋白。结论已初步筛选出HRSV的裂解条件,其对HRSV的裂解效果较好。
OBJECTIVE: To investigate the conditions for the cleavage of human respiratory syncytial virus (HRSV), and to lay the foundation for the development of a purified vaccine against HRSV. Methods Different cleavage agents were used to cleave HRSV at different concentrations and temperatures for different times. The F and G proteins of HRSV were used as target proteins. The proteins were analyzed by Western blot and Western blot. The cleavage effect was verified by electron microscope. Results TritonX-100 at 1% concentration for 90 min at 4 ℃ had better cleavage effect on HRSV than other lysing agents. Electron microscopy showed that the virus samples were cloudy flocculent lysis; Western blot analysis showed that the specific bands of HRSV F protein. Conclusion The conditions for the cleavage of HRSV have been initially screened, which has a better cleavage effect on HRSV.