Construction of recombinant Marek's disease virus (MDV) lacking the meq oncogene and co-expressing AIV-H9N2 HA and NA genes under control of exogenous promoters

文献类型: 外文期刊

第一作者: Zhang, Zhenjie

作者: Zhang, Zhenjie;Chen, Wenqing;Ma, Chengtai;Zhao, Peng;Duan, Luntao;Zhang, Fushou;Su, Hongqin;Li, Sifei;Cui, He;Cui, Zhizhong;Sun, Aijun;Li, Yanpeng

作者机构:

关键词: Marek's disease virus (MDV);H9N2-HA gene;H9N2-NA gene;meq oncogene;Homologous recombination

期刊名称:JOURNAL OF BIOTECHNOLOGY ( 影响因子:3.307; 五年影响因子:3.778 )

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收录情况: SCI

摘要: To develop a recombinant Marek's disease virus (rMDV1) co-expressing the hemagglutinin gene (HA) and neuramidinase gene (NA) from a low pathogenic avian influenza virus (LPAIV) H9N2 strain and lacking the meg oncogene that shares homology with the Jun/Fos family of transcriptional factors, a wild strain of MDV GX0101 was used as parental virus, the HA and NA genes co-expression cassette under control of the CMV and SV40 early promoters was inserted at two meg sites of GX0101 to form a new meg knock-out mutant MDV (MZC12HA/NA) through homologous recombination. MZC12HA/NA was reconstituted by transfection of recombinant BAC-MDV DNA into the secondary chicken embryo fibroblast (CEF) cells. Highly purified MZC12HA/NA was obtained after four rounds of plaque purification and proliferation. In vitro growth properties of recombinant virus were also inspected and concluded that the MZC12HA/NA had the same growth kinetics in CEF cultures as its parental wild type virus GX0101. Southern blot indicated that co-expression cassette was successfully inserted at two copies sites of meg gene, so two meg genes were knocked-out completely. RT-qPCR showed transcription and expression levels of the HA and NA genes were both significantly higher than that of GX0101 own pp38 gene. Indirect fluorescence antibody (IFA) test, and Western blot analyses indicated that HA and NA genes were co-expressed simultaneously under control of the different promoters but meg genes were not. These results herald a new and effective recombinant meg-deleted MDV-based AIV-H9N2 vaccine may be useful in protecting chickens from very virulent MDV and H9N2 challenges

分类号: Q81

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