• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector.
【24h】

Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector.

机译：用杆状病毒载体在昆虫细胞中大肠杆菌β-半乳糖苷酶的强调控表达。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The N-terminal region of the gene encoding polyhedrin, the major occlusion protein of the insect baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV), has been fused to DNA encoding Escherichia coli beta-galactosidase. The fused gene was inserted into the AcNPV DNA genome by cotransfection of insect cells with recombinant plasmid DNA and wild-type AcNPV genomic DNA. Recombinant viruses were selected as blue plaques in the presence of a beta-galactosidase indicator, 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside. Studies of one such virus, L1GP-gal3, indicated that the synthesis of beta-galactosidase is temporally controlled beginning late (20 h) in infection after the release of infectious virus particles from the cell. By 48 h postinfection, a remarkably high level of expression is achieved. On the basis of these results, AcNPV should be a useful vector for the stable propagation and expression of passenger genes in a lepidopteran cell background. A generalized transplacement vector that facilitates the construction and selection of recombinant viruses carrying passenger genes under their own promoter control has also been developed.
机译：编码多角体蛋白的基因的N端区域已被融合到编码大肠杆菌β-半乳糖苷酶的DNA上，该多角体蛋白是昆虫杆状病毒加利福尼亚州产的苜蓿芽孢杆菌核多角体病毒（AcNPV）的主要封闭蛋白。通过将昆虫细胞与重组质粒DNA和野生型AcNPV基因组DNA共转染，将融合基因插入到AcNPV DNA基因组中。在β-半乳糖苷酶指示剂5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷存在下，选择重组病毒作为蓝色噬菌斑。对一种这样的病毒L1GP-gal3的研究表明，β-半乳糖苷酶的合成在感染后期（20小时）从细胞中释放出感染性病毒颗粒后开始受到时间控制。感染后48小时，表达水平显着提高。根据这些结果，AcNPV应该是有用的载体，用于在鳞翅目细胞背景中稳定传播和表达客体基因。还开发了一种通用的移位载体，其有助于构建和选择在其自身的启动子控制下携带客体基因的重组病毒。

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