鸡马立克病疫苗株CVI988/Rispens meq基因编辑及缺失毒株的构建与鉴定

doi:10.11843/j.issn.0366-6964.2020.08.021

Abstract

Abstract: Meq gene is commonly regarded as the main oncogene that plays critical role in the tumorigenesis of Marek's disease (MD). Evolutionarily, there is an obvious sequence difference between the meq genes of vaccine and pathogenic strains with differ virulence. In this study, we have designed the specific guide RNAs (gRNAs) targeting to the meq gene of the vaccine strain CVI988/Rispens, to make the plasmid pX459-gRNA. The CEF cells were transfected with pX459-gRNA plasmids and were infected with CVI988/Rispens virus to produce meq-gene mutated viral particles. Utilizing the CRISPR/Cas9-based gene editing technique, together with the viral plaque purification, PCR analysis, DNA sequencing and indirect immunofluorescence assay (IFA), we successfully generated a meq-deleted mutant named as CVI988Δmeq-C7. Our data provides an important basis for further screening of the monoclonal antibodies against MD vaccine MEQ protein and the development of differential diagnostic reagents.

Key words: Marek's disease, CVI988/Rispens, meq gene, CRISPR/Cas9, gene editing

CLC Number:

S852.659.1

YANG Sen, TENG Man, LIU Jinling, ZHOU Ziyu, ZHENG Luping, CHU Yushu, DING Ke, YU Zuhua, LUO Jun. Construction of meq Deleted strain by Gene Editing of Marek's Disease Vaccine Strain CVI988/Rispens via the CRISPR/Cas9 System and Identification[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1970-1976.

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Construction of meq Deleted strain by Gene Editing of Marek's Disease Vaccine Strain CVI988/Rispens via the CRISPR/Cas9 System and Identification

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