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Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus

  • RESEARCH ARTICLE
  • Published:
Virologica Sinica

Abstract

Baculoviruses have been widely used as a vector for expressing foreign genes. Among numerous baculoviruses, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most frequently used and it encodes 155 open reading frames (ORFs). Here, we systematically investigated the impact of 42 genes of AcMNPV on the production of infectious budded viruses (BVs) by constructing gene-knockout bacmids and subsequently conducting transfection and infection assays. The results showed that among the 39 functionally unverified genes and 3 recently reported genes, 36 are dispensable for infectious BV production, as the one-step growth curves of the gene-knockout viruses were not significantly different from those of the parental virus. Three genes (ac62, ac82 and ac106/107) are essential for infectious BV production, as deletions thereof resulted in complete loss of infectivity while the repaired viruses showed no significant difference in comparison to the parental virus. In addition, three genes (ac13, ac51 and ac120) are important but not essential for infectious BV production, as gene-knockout viruses produced significantly lower BV levels than that of the parental virus or repaired viruses. We then grouped the 155 AcMNPV genes into three categories (Dispensable, Essential, or Important for infectious BV production). Based on our results and previous publications, we constructed a schematic diagram of a potential mini-genome of AcMNPV, which contains only essential and important genes. The results shed light on our understanding of functional genomics of baculoviruses and provide fundamental information for future engineering of baculovirus expression system.

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Acknowledgements

This research was supported by the grants from the National Natural Science Foundation of China (No. 31872640) and the Key Research Program of Frontier Sciences of the Chinese Academy of Sciences (grant no. QYZDJ-SSW-SMC021), and the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB11030400). The authors acknowledge colleagues from the Core Facility and Technical Support of the Wuhan Institute of Virology for technical assistance and we would like to thank Dr. Basil M. Arif for kindly editing the manuscript.

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  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, 430071, China

    Tong Chen, Xiaoyan Duan, Hengrui Hu, Yu Shang, Yangbo Hu, Fei Deng, Hualin Wang, Manli Wang & Zhihong Hu

  2. University of Chinese Academy of Sciences, Beijing, 10049, China

    Tong Chen, Xiaoyan Duan & Hengrui Hu

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  1. Tong Chen
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TC, MW and ZH designed the experiments. TC and XD performed the experiments. TC and HH analyzed the data. TC, MW and ZH wrote the manuscript. YS, YH, FD, HW, MW and ZH edited and commented on the manuscript.

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Correspondence to Manli Wang or Zhihong Hu.

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Chen, T., Duan, X., Hu, H. et al. Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus. Virol. Sin. 36, 762–773 (2021). https://doi.org/10.1007/s12250-021-00355-1

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