• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biotechnology Progress >Enhancement and Prolongation of Baculovirus-Mediated Expression in Mammalian Cells:Focuses on Strategic Infection and Feeding
【24h】

Enhancement and Prolongation of Baculovirus-Mediated Expression in Mammalian Cells:Focuses on Strategic Infection and Feeding

机译：增强和延长杆状病毒介导的哺乳动物细胞中的表达：战略感染和喂养的重点。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The baculovirus/insect cell system has been widely used for recombinant protein production.Since the finding that baculovirus as a gene delivery vehicle into mannalian cells ahve been reported.In this study,we intended to explore the possibility of utilizing a baculovirus/mammalina cell system as a nonlytic continuous protein production system.Arecombinant baculovirus vector carrying enhaneced green fluorescent protein (EGFP)under the control of cytomegalovirus immediate-early (CMV-IE) promoter was constructed.This virus was used to infedt four common mannalian cell lines and HeLa was found to yield the highest experession level.Additions of butyrate and valproic acid both enhanced the expression level Addition of butyrate and valproic acid both enhanced the expression levelbut butyrate exhibited a more profound effect.More importantly HeLa cells were found to be superinfected gy baculovirus,a result not observed in the vonventional baculovirus/insect cell system.The effects of multiplicity of infection (MOI) and infection eiming were also compared High MOI up to 800 increased the expression in the short term (4 days),but the relatively higher cell death and lower cell density compromised the overall protein yield therafter.The highest overall expression for a long term was obtained at MOI=200 when the cells were initally infected at the mid-exponential phase and superinfected with additional baculovirus (MOI=200) together with a one-time supplement of butyrate.In summary the strategic infection and feeding enhanced the expression level9-fold (compared with unsuperinfected culture) adn prolonged the duration of expression to 16 days.This study reveals that thsi baculovirus/mammalian cell system has great potential to become a novel continuons nonlytic expression system.
机译：杆状病毒/昆虫细胞系统已被广泛用于重组蛋白的生产。自从发现杆状病毒作为基因传递载体进入曼纳期细胞以来，这项研究就一直在探索利用杆状病毒/哺乳动物细胞系统的可能性。构建了带有增强型绿色荧光蛋白（EGFP）的重组杆状病毒载体，该载体在巨细胞病毒即早启动子（CMV-IE）启动子的控制下进行了构建，该病毒被用于侵染四种常见的曼纳期细胞系，HeLa丁酸酯和丙戊酸的添加均能提高表达水平丁酸酯和丙戊酸的添加均能提高表达水平但丁酸酯显示出更深远的作用。更重要的是，发现HeLa细胞是超级感染的杆状病毒，a在典型的杆状病毒/昆虫细胞系统中未观察到结果。还比较了感染的感染性（MOI）和感染开始时间，短期内（4天）高达800的高MOI会增加表达，但相对较高的细胞死亡和较低的细胞密度会损害随后的总蛋白产量。当细胞在指数中期被初始感染并用额外的杆状病毒（MOI = 200）和丁酸的一次性补充物进行超级感染时，获得了长期的MOI = 200。表达水平的9倍（与未感染的培养物相比）和表达持续时间延长至16天。这项研究表明杆状病毒/哺乳动物细胞系统具有成为新的连续性非裂解表达系统的巨大潜力。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号