Airenne Kari J, Peltomaa Erik, Hytönen Vesa P, Laitinen Olli H, Ylä-Herttuala Seppo
AI Virtanen Institute, Department of Molecular Medicine and Biotechnology, University of Kuopio and Kuopio University Hospital, Kuopio, Finland.
Nucleic Acids Res. 2003 Sep 1;31(17):e101. doi: 10.1093/nar/gng102.
An improved method for the generation of recombinant baculoviruses by Tn7-mediated transposition is described. The method is based on the modified donor vector (pBVboost) and an improved selection scheme of the baculovirus bacmids in Escherichia coli with a mutated SacB gene. Recombinant bacmids can be generated at a frequency of approximately 10(7)/microg of donor vector with a negligible background. This easy-to-use and efficient pBVboost system provides the basis for a high-throughput generation of recombinant baculoviruses as well as a more convenient way to produce single viruses. The introduced selection scheme is also useful for the construction of other vectors by transposition in E.coli.
本文描述了一种通过Tn7介导的转座产生重组杆状病毒的改进方法。该方法基于修饰的供体载体（pBVboost）以及在大肠杆菌中对带有突变SacB基因的杆状病毒杆粒的改进筛选方案。重组杆粒可以以大约10(7)/μg供体载体的频率产生，背景可忽略不计。这个易于使用且高效的pBVboost系统为高通量产生重组杆状病毒提供了基础，同时也为生产单一病毒提供了更便捷的方法。引入的筛选方案对于通过在大肠杆菌中转座构建其他载体也很有用。
