EXPRESSION AND RECOVERY OF RECOMBINANT ROTAVIRUS VP4 FROM INSECT CELL CULTURES

从昆虫细胞培养物中表达和回收重组轮状病毒 VP4

• 批准号: 3809744
• 负责人: M GORZIGLIA
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3809744
• 关键词: Insecta; Rotavirus; genetic manipulation; insect virus; protein purification; tissue /cell culture; virus protein

The possibility of producing and purifying large quantities of recombinant rotavirus VP4 from insect cell cultures was investigated. Spodoptera frugiperda (Sf9) cells were grown in large spinner flasks and infected with a baculovirus expressing the rotavirus OSU VP4 gene. The expressed VP4 subsequently was purified from such cultures to determine the yield of recombinant protein. Conditions for growth of cells and high yield of VP4 were optimized to improve the expression of VP4. Also, a practical purification scheme was developed to maximize the yield of VP4 from insect cell cultures and to reduce cost and complexity of a potential industrial-scale purification process.

生产和纯化大量重组蛋白的可能性 研究了来自昆虫细胞培养物的轮状病毒VP4。夜蛾 在大的转瓶中培养草地贪夜蛾（Sf9）细胞，并感染 表达轮状病毒OSU VP4基因的杆状病毒。表达的VP 4 随后从这样的培养物中纯化，以确定 重组蛋白细胞生长和VP4高产率的条件 优化以提高VP4的表达。此外，一个实用的 本研究建立了一种从昆虫中最大限度地获得VP4的纯化方案 细胞培养和降低成本和复杂性的潜力 工业规模的纯化工艺。