一种用于蛋白质水平研究的感染性登革病毒纯化的无梯度方法。
A gradient-free method for the purification of infective dengue virus for protein-level investigations.
作者信息
Jensen Stephanie M, Nguyen Celina T, Jewett John C
机构信息
The Department of Chemistry and Biochemistry, The University of Arizona, 1306 E. University Blvd., Tucson, AZ 85721, USA.
The Department of Chemistry and Biochemistry, The University of Arizona, 1306 E. University Blvd., Tucson, AZ 85721, USA.
出版信息
J Virol Methods. 2016 Sep;235:125-130. doi: 10.1016/j.jviromet.2016.05.017. Epub 2016 Jun 2.
Dengue virus (DENV) is a mosquito-transmitted flavivirus that infects approximately 100 million people annually. Multi-day protocols for purification of DENV reduce the infective titer due to viral sensitivity to both temperature and pH. Herein we describe a 5-h protocol for the purification of all DENV serotypes, utilizing traditional gradient-free ultracentrifugation followed by selective virion precipitation. This protocol allows for the separation of DENV from contaminating proteins - including intact C6/36 densovirus, for the production of infective virus at high concentration for protein-level analysis.
登革病毒（DENV）是一种通过蚊子传播的黄病毒，每年感染约1亿人。由于病毒对温度和pH值敏感，用于纯化登革病毒的多日方案会降低感染滴度。在此，我们描述了一种5小时的方案，用于纯化所有登革病毒血清型，该方案采用传统的无梯度超速离心，随后进行选择性病毒粒子沉淀。该方案能够从包括完整的C6/36浓病毒在内的污染蛋白中分离出登革病毒，以高浓度生产感染性病毒用于蛋白质水平分析。
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