文献类型：期刊文献

英文题名：Genome-wide nonessential gene identification of Autographa californica multiple nucleopolyhedrovirus

作者：Yu, Yue[1];Zhang, Tong[1];Lu, Dongbo[1];Wang, Jing[1];Xu, Zhenhe[1];Zhang, Yuanxing[2];Liu, Qin[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2023

卷号：863

外文期刊名：GENE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000992679700001)】；

基金：This work was supported by Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism grant 2021 Sci & Tech 03-28 (Shanghai Municipal Education Commission) and The Fundamental Research Funds for the Central Universities (222201211716).

语种：英文

外文关键词：Autographa californica multiple; nucleopolyhedrovirus (AcMNPV); Baculovirus; Gene deletion; Insect cell; Minimal genome

摘要：The Baculovirus Expression Vector System (BEVS) is an insect cell-based heterologous protein expression system that possesses powerful potential in the development of protein drugs and vaccines. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most widely-used vector in BEVS with 151 open reading frames (ORFs) containing essential and nonessential genes. Deletion of nonessential genes has many advantages including increased foreign gene insertion. In this study, the lambda red recombination system was used to knock out genes in a modified AcMNPV that carried an enhanced yellow fluorescent protein (eYFP) at the Ac126-Ac127 locus. Eighty genes were almost completely deleted respectively and 69 gene knockout AcMNPVs (KOVs) were obtained to evaluate their infection efficiency. After infecting Spodoptera frugiperda 9 (Sf9) cells, 51 KOVs including 62 genes showed similar infectivity as wide type (WT) and hence were defined as nonessential genes. However, 18 KOVs produced fewer infectious virions, indicating that these genes were influential in the pro-duction of progeny viruses. Combining our research with previous studies, a desired minimal AcMNPV genome containing 86 ORFs and all of the homologous regions (hrs) was brought up, facilitating genetic modification of baculovirus vectors and improvement of recombinant protein expression in the future.