条件性敲除D1133L基因的重组非洲猪瘟病毒的构建及增殖特性

doi:10.11843/j.issn.0366-6964.2023.02.027

Abstract

Abstract: Previous studies have shown that the D1133L gene encoded by African swine fever virus (ASFV) is crucial to ASFV replication. In order to further explore the role of D1133L in ASFV replication, in this study, we used homologous recombination technology and Escherichia coli lac repression operating system to knockout D1133L gene conditionally, constructed recombinant transfer vector ASFV Δi130 using pUC118 as vector, transfected BMDM cells with recombinant transfer vector, and infected BMDM cells with ASFV CN/GS/2018 wild type strain. In the presence of isopropyl-β-D-thiogalactoside (IPTG), conditional knockout D1133L recombinant strain vD1133Li was identified by green fluorescence and qPCR. The replication of the recombinant virus strain and the parent virus strain in PAM cells was observed with fluorescence microscope. Differences between vD1133Li and the parent virus strain were analyzed by qPCR. The replication difference between conditionally knockout D1133L recombinant strain with or without IPTG was also further investigated. The vD1133Li with conditional deletion of D1133L was constructed successfully in this study. Replication level of vD1133Li was significantly lower than that of the wild type virus strain. In MA-104 cell lines that overexpressing stably D1133L, the replication ability of vD1133Li was recovered. In summary, D1133L is crucial for ASFV replication. These results provide a basis for further research on the function of D1133L in ASFV replication, the pathogenic mechanism and the development of D1133L targeted drugs.

Key words: African swine fever virus, conditional D1133L gene knockout recombinant virus, proliferation characterizations

CLC Number:

S852.659.1

ZHANG Ting, FENG Tao, YANG Jinke, HAO Yu, YANG Xing, ZHANG Dajun, SHI Xijuan, YAN Wenqian, CHEN Lingling, LIU Xiangtao, ZHENG Haixue, ZHANG Keshan. Construction and Growth Characteristics of Recombinant African Swine Fever Virus with Conditional Deletion of D1133L Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 706-714.

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Construction and Growth Characteristics of Recombinant African Swine Fever Virus with Conditional Deletion of D1133L Gene

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