In the Antheraea pernyi multicapsid nucleopolyhedrovirus (AnpeNPV)-based expression vector system, the frequency of homologous recombination events between wild-type AnpeNPV DNA and the transfer vector is low, resulting in a small amount of recombinant virus. Previous reports have indicated that linearized baculovirus DNA can increase the proportion of recombinant virus relative to the total progeny. To improve the recombination efficiency, we constructed a linearized derivative of AnpeNPV, referred to as AnpeNPV^PhEGFP-AvrII, in which egfp flanked by AvrII restriction sites was located at the polyhedrin locus and driven by the polyhedrin promoter. Linear AnpeNPV DNA was obtained by the treatment of AnpeNPV^PhEGFP-AvrII genomic DNA with AvrII endonuclease. The infectivity and recombinogenic activity between the linearized and circular viral DNA were evaluated by quantitative real-time polymerase chain reactions. We demonstrated that the linearized AnpeNPV DNA produced only small numbers of infectious budded viruses, accounting for approximately 4.5% of the budded virus production of wild-type AnpeNPV DNA in A. pernyi pupae. However, the linearized AnpeNPV DNA substantially increased recombinant virus production after cotransfection with an appropriate transfer vector; relative abundance of the recombinant virus was approximately 5.5-fold higher than that of the wild-type AnpeNPV DNA in A. pernyi pupae. The linearization of AnpeNPV DNA will facilitate the purification of recombinant viruses using the AnpeNPV-based expression vector system and the construction of an AnpeNPV-based bacmid system.

中文翻译：

---

杆状病毒衍生物的生产，以产生线性化的An蚕（鳞翅目：Saturniidae）多衣壳核多角体病毒基因组DNA。

在柞蚕multicapsid核型（AnpeNPV）系的表达载体系统，野生型DNA AnpeNPV和转移载体之间的同源重组事件的频率是低的，从而产生重组病毒的量小。以前的报道表明线性化的杆状病毒DNA可以增加重组病毒相对于总子代的比例。为了提高复合效率，我们构建了线性化的衍生物AnpeNPV的，被称为AnpeNPV ^{PhEGFP -Avr} II，其中EGFP侧翼通过的Avr II限制性位点被定位在多角体蛋白基因座和由多角体蛋白启动子驱动。通过处理AnpeNPV获得线性AnpeNPV DNA^{PhEGFP -Avr} II与基因组DNA的Avr II内切酶。通过定量实时聚合酶链反应评估线性和环状病毒DNA之间的感染性和重组活性。我们证明了线性AnpeNPV DNA只产生的感染性病毒发芽小的数字，占出芽病毒生产野生型AnpeNPV DNA的约4.5％柞蚕蛹。然而，在用合适的转移载体共转染后，线性化的AnpeNPV DNA大大增加了重组病毒的产生；重组病毒的相对丰度比A. pernyi中的野生型AnpeNPV DNA高约5.5倍蛹。AnpeNPV DNA的线性化将有助于使用基于AnpeNPV的表达载体系统和基于AnpeNPV的杆粒系统构建重组病毒。