Construction and Characterization of a Novel Bacmid AcBac-Syn Based on a Synthesized Baculovirus Genome  

作　　者：Yu Shang Hengrui Hu Xi Wang Hualin Wang Fei Deng Manli Wang Zhihong Hu 

机构地区：[1]Key Laboratory of Prevention and Control Agents for Animal Bacteriosis(Ministry of Agriculture and Rural Affairs)&Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology,Institute of Animal Husbandry and Veterinary,Hubei Academy of Agricultural Sciences,Wuhan 430064,China [2]State Key Laboratory of Virology,Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China [3]University of Chinese Academy of Sciences,Beijing 100049,China

出　　处：《Virologica Sinica》2021年第6期1566-1574,共9页中国病毒学（英文版）

基　　金：This work was supported by the grants from the National Science Foundation of China(Grant Nos.31800143 and 31872640);the Key Research Program of Frontier Sciences of the Chinese Academy of Sciences(Grant No.QYZDJ-SSW-SMC021);the Hubei Provincial Innovation Center of Agricultural Sciences and Technology(Grant No.2019-620-000-001-017)。

摘　　要：Baculoviruses are large DNA viruses which have been widely used as expression vectors and biological insecticides.Homologous recombination and Bac-to-Bac system have been the main methods for manipulating the baculovirus genome.Recently, we generated a synthetic baculovirus Ac MNPV-WIV-Syn1 which fully resembled its parental virus Autographa californica multiple nucleopolyhedrovirus(Ac MNPV). Here, we report the modification of Ac MNPV-WIV-Syn1 into a novel bacmid, Ac Bac-Syn, which can be used as a backbone for Bac-to-Bac system. To achieve this, a vector contained a Lac Z:att Tn7 and egfp cassette was constructed, and recombined with a linearized Ac MNPV-WIV-Syn1 genome by transformation-associated recombination in yeast to generate bacmid Ac Bac-Syn. The bacmid was then transfected to insect cells and the rescued virus showed similar biological characteristics to the wild-type virus in terms of the kinetics of budded virus production, the morphology of occlusion bodies, and the oral infectivity in insect larvae. For demonstration, a red fluorescent protein gene Dsred was transposed into the att Tn7 site by conventional Bac-to-Bac method, and the transfection and infection assays showed that Ac Bac-Syn can be readily used for foreign gene insertion and expression.Ac Bac-Syn has several advantages over the conventional Ac MNPV bacmids, such as it contains an egfp reporter gene which facilitates visualization of virus propagation and titration;its DNA copy numbers could be induced to a higher level in E. coli;and the retaining of the native polyhedrin gene in the genome making it an attractive system for studying the functions of gene related to occlusion body assembly and oral infection.

关 键 词：Synthetic genome BACULOVIRUS Autographa californica multiple nucleopolyhedrovirus(AcMNPV) Large DNA virus 

分 类 号：Q939.4[生物学—微生物学]