...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Current Science: A Fortnightly Journal of Research >Development of a rapid and efficient BmNPV baculovirus expression system for application in mulberry silkworm, Bombyx mori
【24h】

Development of a rapid and efficient BmNPV baculovirus expression system for application in mulberry silkworm, Bombyx mori

机译：快速高效的BmNPV杆状病毒表达系统的开发，用于桑蚕Bombyx mori

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

Silkworm–baculovirus gene expression system is one of the most powerful eukaryotic expression systems. However, due to very low recombination frequency, the traditional method to construct and obtain pure recombinant baculovirus requires plaque assaythat is time-consuming and also utilizes skillful techniques. In order to overcome this disadvantage, a rapid BmNPV expression system applicable to silkworm was constructed based on the working principle of AcMNPV Bac-to-Bac system. A large 8.6 kb fragment containing the low-copy-number mini-F replicon, a kanamycin resistance marker and a segment of DNA encoding the lacZ a peptide from the AcMNPV bacmid, was cloned into polyhedrin locus of BmNPV genome to replace the polyhedrin gene. This recombinant,designated as BmBacmid, was transformed into Escherichia coli DH10 b strain, in which a helper plasmid encoding the transposase was already transformed. We designated the DH10 b strain containing BmBacmid and helper as DH10BmBac. With this bacterium, therecombinant baculovirus can be rapidly and easily generated through gene transposition. This system has an advantage of high recombination frequency, simple manipulation, high-efficiency and is time-saving. This approach permits large potential value inrecombinant protein production using silkworm as a ‘biofactory’ in the future biotechnological industry and can become a powerful tool for structural and functional analysis of protein in post-genomic era.
机译：蚕杆状病毒基因表达系统是最强大的真核表达系统之一。然而，由于非常低的重组频率，构建和获得纯重组杆状病毒的传统方法需要噬菌斑测定，这是费时的并且还需要熟练的技术。为了克服这一缺点，基于AcMNPV Bac-to-Bac系统的工作原理，构建了适用于家蚕的快速BmNPV表达系统。将一个8.6 kb的大片段克隆到BmNPV基因组的多面体基因座中，以取代多面体基因，该片段包含低拷贝数的mini-F复制子，卡那霉素抗性标记和一段编码来自AcMNPV杆粒的lacZ a肽的lac DNA片段。将该重组体命名为BmBacmid，将其转化到大肠杆菌DH10b菌株中，其中编码转座酶的辅助质粒已经被转化。我们将含有BmBacmid和辅助分子的DH10b菌株命名为DH10BmBac。利用这种细菌，可以通过基因转座快速而容易地产生重组杆状病毒。该系统具有重组频率高，操作简单，效率高且节省时间的优点。这种方法可以在未来的生物技术产业中使用家蚕作为“生物工厂”来生产具有巨大潜在价值的重组蛋白，并且可以成为后基因组时代蛋白质结构和功能分析的强大工具。

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