Construction of a recombinant MDV vector vaccine integrating the NDV-F gene
In order to prevent co-infection of Marek's disease virus(MDV)and Newcastle disease virus(NDV)in clinic,this study used bacterial artificial chromosome(BAC)as the gene editing platform to integrate the F gene of NDV into the genome of MDV double-gene deletion strain Md5 BAC ΔmeqΔLorf9 through two-step Red-mediated homologous recombination technology.The ex-pression of Ⅰ-SceⅠ enzyme was induced to knocked out the kanamycin resistance gene,and the recombinant live vector vaccine candidate strain Md5 BACΔmeqΔLorf9-F was constructed.PCR and restriction fragment polymorphism(RFLP)assays confirmed the integrity and correctness of MDV BAC clones containing the F gene.Then,the recombinant plasmid was transfected into chicken embryo fibroblasts to obtain recombinant virus,which was identified by indirect immuno-fluorescence assay.The results of plaque size determination and in vitro growth curve showed that the insertion of F gene did not affect viral replication in vitro.Meanwhile,IFA test confirmed the expression of F protein.Overall,these results indicate that we successfully construct recombinant MDV expressing NDV protective antigen F gene,which provides basis for the prevention and con-trol of NDV and MDV co-infection.
NETL
NSTL
万方数据
