Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus
Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus
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对 42 个苜蓿银纹夜蛾多核多角体病毒基因的系统分析鉴定出另外 6 个参与传染性芽生病毒产生的基因
DOI：
10.1007/s12250-021-00355-1
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发表时间：
2021-03-08
影响因子：
5.5
通讯作者：
Hu, Zhihong
中科院分区：
文献类型：
--
作者：
Chen, Tong;Duan, Xiaoyan;Hu, Zhihong
杆状病毒已被广泛用作表达外源基因的载体。在众多杆状病毒中，苜蓿银纹夜蛾多核多角体病毒（AcMNPV）是最常用的，它编码155个开放阅读框（ORF）。在这里，我们通过构建基因敲除杆粒并随后进行转染和感染测定，系统地研究了 AcMNPV 的 42 个基因对传染性芽生病毒 (BV) 产生的影响。结果显示，在39个功能未经验证的基因和3个最近报道的基因中，有36个基因对于感染性BV的产生是可有可无的，因为基因敲除病毒的一步生长曲线与亲本病毒没有显着差异。三个基因（ac62、ac82和ac106/107）对于感染性BV的产生至关重要，因为其缺失导致感染性完全丧失，而修复后的病毒与亲本病毒相比没有显着差异。此外，三个基因（ac13、ac51和ac120）对于传染性BV的产生很重要，但不是必需的，因为基因敲除病毒产生的BV水平明显低于亲本病毒或修复病毒。然后，我们将 155 个 AcMNPV 基因分为三类（可有可无、必需或对于传染性 BV 产生很重要）。根据我们的结果和之前的出版物，我们构建了 AcMNPV 潜在迷你基因组的示意图，其中仅包含必需和重要的基因。这些结果阐明了我们对杆状病毒功能基因组学的理解，并为未来杆状病毒表达系统的工程提供了基础信息。
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,ac82andac106/107) 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,ac51andac120) 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.