表达禽网状内皮组织增生病病毒env基因的重组马立克病毒的生物学特性

doi:10.11843/j.issn.0366-6964.2018.07.018

Abstract

Abstract:

The study aimed to construct a Marek's disease virus(MDV) recombinant expressing the env gene of Reticuloendotheliosis virus(REV), and explore its biocharacteristics preliminarily. Firstly, env gene was amplified from the DNA of REV SNV strain and cloned into eukaryotic expression vector pcDNA3.1(+). The env gene expression cassette and the kanamycin resistance gene(kan^r) amplified from the recombinant plasmid were then cloned into pUC18 vector, respectively. Using the primers containing homologous arms on both sides of the meq gene of MDV strain SC9-1, the REV env gene expression cassette and the kan^r gene were amplified and integrated into the site of meq gene in the genome via Red E/T homologous recombination system. The kan^r gene was further knocked out by the Arab sugar induced flg recombinant enzyme to obtain the recombinant BAC plasmid, naming SC9-env BAC. The recombinant MDV strain SC9-env was rescued after transfecting the SC9-env BAC plasmid into CEF cells, and identified by indirect immunofluorescence(IFA) using the MDV monoclonal antibody H19 and the REV monoclonal antibody 11B118. Replication of SC9-env on CEF cells, the pathogenicity of SC9-env, and the protective efficacy of SC9-env against challenge of MDV and REV in SPF chickens were further evaluated. The results showed that the REV env protein was stably expressed by the recombinant virus SC9-env. The recombinant virus SC9-env showed similar replication capacity to the parental virus SC9-1, but had no pathogenicity to SPF chickens. In addition, SC9-env protected 92% of SPF chickens against challenge of MDV strain rMd5, which showed no significant difference with SC9-1, and eased the weight loss and the decreased inactivated antibody levels caused by REV infection. In conclusion, the constructed MDV recombinant expressing the env gene of REV possesses good protective efficacy against infection of MDV and REV in SPF chickens.

Key words: reticuloendotheliosis virus, env gene, homologous recombination, Marek's disease virus, biocharacteristics

CLC Number:

S858.315.3

ZHOU Zhong-wen, LIN Gui-fang, WANG Hao-hao, SU Shuai, CUI Zhi-zhong, SUN Shu-hong. Biocharacteristics of Recombinant Marek's Disease Virus Expressing the env Gene of Reticuloendotheliosis Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(7): 1482-1491.

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Biocharacteristics of Recombinant Marek's Disease Virus Expressing the env Gene of Reticuloendotheliosis Virus

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