Bac-to-Bac系统在杆状病毒功能基因研究上的应用

Study of the Functional Baculovirus Genes Using Bac-to-Bac System

利用Bac-to-Bac系统,在大肠细菌中复制增殖杆状病毒质粒bacmid,并通过RecA介导法、ET-recombination法在bacmid DNA上缺失或插入功能基因,构建功能基因缺陷型或补回型的杆状病毒质粒。该质粒转染到昆虫细胞中产生重组病毒,进而在细胞甚至虫体水平上研究基因的功能,极大改善了传统上用空斑实验筛选重组病毒的不足。

The functional gene was deleted or inserted through homologous recombination from an baculovirus genome propagated as a bacmid DNA in E. coli, generating a me53（gene knockout or repair bacmid. The knockout or repair bacmid was transfected into insect cells to generate a recombination baculovirus. It was available to study viral gene function through transfected cell or infected insect and improve the limitation of the traditional plaques formation test producing recombination baculovirus.