Platform establishment of the Cre-loxP recombination system for genetic manipulation of the Lumpy skin disease virus

文献类型： 外文期刊

第一作者： Ren, Shanhui

作者： Ren, Shanhui;Zhang, Yuzhe;Wang, Xiangwei;Wang, Shasha;Sun, Yuefeng;Yin, Xiangping;Chen, Haotai;Gao, Xiaolong;Tong, Lina

作者机构：

关键词： Lumpy skin disease virus; Cre-loxP system; Recombinant virus; MDBK-Cre cell line

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：3.3； 五年影响因子：3.5 ）

ISSN： 0378-1135

年卷期： 2024 年 294 卷

页码：

收录情况： SCI

摘要： Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in Asia, including China. Genetic manipulation of the LSDV is essential for the elucidation of the pathogenic mechanism and biological function of the LSDVencoded protein. In this study, we established a platform for the Cre-loxP recombination system under a modified early-late H5 promoter of the VACV for quick construction of the recombinant LSDV virus. The recombinant virus, LSDV-EGFP-Delta TK, was purified and obtained using serial limited dilution and picking the single cells methods. Using the lentiviral package system, a Cre recombinase enzyme stable expression MDBK cell line was established to supply the Cre recombinase for the reporter gene excision. A genetically stable, safe TK genedeleted LSDV (LSDV-Delta TK) was constructed using homologous recombination and the Cre-loxP system. It was purified using limited dilution in the MDBK-Cre cell line. Establishing the Cre-loxP recombination system will enable sequential deletion of the interested genes from the LSDV genome and genetic manipulation of the LSDV genome, providing technical support and a platform for developing the attenuated LSDV vaccine.

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