Construction And Expression Of Recombinant Marek's Disease Virus CVI988 Strain Transferring Plasmid Vector With F Gene Of Newcastle Disease Virus

Newcastle disease is an acute infectious disease caused by Newcastle disease virus in chicken andturkey. Newcastle disease' vaccination with live virus has been used since 1940s,which lead torespiratory complications and cause bacterium infection. These vaccines were used many times toproduce immunity protection in chicken and their immunes were disturbed by mother' antibody inchicken .So we construct recombinant virus which express NDV antigen gene to deal with theseproblem. Marek's Disease is a highly contiguity infectivity tumour disease caused by Marek's Disease Virusin chicken. The MDV vaccine viruses are considered one of the most potent vectors for polyvalent livevaccines expressing foreign antigens related to vaccine-induced immunity against poultry diseases.There are great homologue and similarity between MDV 1 type and vvMDV,so MDV1 can preventMD efficiently. Considering polyvalent recombinant vaccine with much advantage to prevent poultry diseasesefficiently,we try to construct recombinant MDV expressing NDV F gene controlled by MDVglycoprotein B promoter .we hope this recombinant virus can avoid virus rejecting to foreign promoterand avoid disturb of mother' antibody to prevent NDV and vvMDV efficiently, One pair of primers were synthesized according fusion protein gene of NDVF48E9 strainnucleotide sequence published on GenBank The NDV F48E9 strain Fusion Protein gene was amplifiedand cloned into the multi clone site of eukaryotic expression vector pIRES to form pIRESF. One pair ofthe primers were synthesized according vector pIRES gene nucleotide sequence published onGenBank .This pair of primers were used to amplify 2900bp DNA fragment containing F gene with IVSand polyA and was cloned into multi clone site of vector pBluescprint II+SK with gB promoter of MDV.At last ,the DNVF gene expression cassette was inserted into us10 gene to give rise to the transferringvector pUS10F. We choose CVI988 strain as virus carrier to construct recombinant virus, because the immunityeffect of CVI988 strain is better than HVT, at the same time, there are great homologue and similaritybetween antigen of MDVI and vvMDV .In theory,CVI988 strain has better pertinence immunityresponse reaction ,especially for vvMDV and vv+MDV. In order to improve expression level ofrecombinant virus ,the transferring vector was transfected 293T cells and the specific fluorescent andresult of PCR in the transfected 293T cells was observed. The result is the basis of obtainingrecombinant MDVCVI988 expressing NDV F gene.