摘要
The production of a recombinant baculovirus expression vector normally involves mixing infectious virus DNA with a plasmid-based transfer vector and then co-transfecting insect cells to initiate virus infection. The aim of this chapter is to provide an update on the range of baculovirus transfer vectors currently available. Some of the original transfer vectors developed are now difficult to obtain but generally have been replaced by superior reagents. We focus on those that are available commercially and should be easy to locate. These vectors permit the insertion of single or multiple genes for expression, or the production of proteins with specific peptide tags that aid subsequent protein purification. Others have signal peptide coding regions permitting protein secretion or plasma membrane localization. A table listing the transfer vectors also includes information on the parental virus that should be used with each one. Methods are described for the direct insertion of a recombinant gene into the virus genome without the requirement for a transfer vector. The information provided should enable new users of the system to choose those reagents most suitable for their purposes.
摘要译文
重组杆状病毒表达载体通常涉及将感染性病毒DNA与基于质粒的转移载体混合，然后共转染昆虫细胞以启动病毒感染本章的目的是提供目前可用的杆状病毒转移载体的范围的更新开发的一些原始转移载体现在难以获得，但通常已被优选的试剂替代我们专注于商业上可用的并且易于定位。这些载体允许插入单个或多个基因用于表达，或产生具有特异性肽标签的蛋白质，有助于随后的蛋白质纯化其他具有允许蛋白质分泌或质膜定位的信号肽编码区列出转移载体的表格还包括有关每一个应使用的亲代病毒的信息描述了将重组基因直接插入到病毒基因组中而不需要转移载体的方法所提供的信息应使该系统的新用户能够选择最适合其用途的那些试剂
Robert D. Possee (3); (4); Linda A. King (5);. Baculovirus Transfer Vectors. Baculovirus and Insect Cell Expression Protocols[M].DE: Springer, 2016: 51-71