Construction of a Baculovirus Derivative to Produce Linearized Antheraea pernyi (Lepidoptera: Saturniidae) Multicapsid Nucleopolyhedrovirus Genomic DNA

文献类型： 外文期刊

第一作者： Zhao, Zhenjun

作者： Zhao, Zhenjun;Ye, Bo;Yue, Dongmei;Li, Peipei;Zhang, Bo;Wang, Linmei;Fan, Qi

作者机构：

关键词： Antheraea pernyi; nucleopolyhedrovirus; linearization; homologous recombination; quantitative real-time polymerase chain reaction

期刊名称：JOURNAL OF INSECT SCIENCE （ 影响因子：1.857； 五年影响因子：1.904 ）

ISSN：

年卷期： 2020 年 20 卷 2 期

页码：

收录情况： SCI

摘要： In the Antheraea pemyi 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 )-Avrll, in which egfp flanked by AvAl 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)-Avrll genomic DNA with AvAl 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. pemyi 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. pemyi 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.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序