Chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and construction method of candidate strain
A technology of Newcastle disease virus and maternal antibody is applied in the field of chimeric Newcastle disease virus vector H9 live vaccine candidate strain that overcomes the influence of Newcastle disease maternal antibody in chicks and its construction, and can solve problems such as restricting the application of NDV vector vaccine.
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Embodiment 1
[0033] Example 1: Construction of recombinant plasmid pNDV / rAI4-T4FHN
[0034] Construction Step 1: Modification of Newcastle Disease Virus AI4 Genome
[0035] Biomaterial preparation:
[0036] The full-length expression vector pNDV / rAI4 of NDV AI4 strain (Hu Z, Hu S, Meng C, et al. Generation of a Genotype VII Newcastle Disease Virus Vaccine Candidate with High Yield in Embryonated Chicken Eggs. Avian Diseases 2011,55(3):1759- 1769) was constructed and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases of the Ministry of Agriculture of Yangzhou University.
[0037] pCR2.1 vector: purchased from Invitrogen; AMV reverse transcriptase, High fidelity DNApolymerase, T4DNA ligase, and Agarose Gel DNA Extraction Kit were purchased from Roche; transfection reagent SuperFect and plasmid extraction kit (QIAprep Spin MiniPrep Kit) were The products of QIAGEN; the rest of the conventional reagents were of domestic analytical grade.
[0038] Design mutati...
Embodiment 2
[0078] The construction model of recombinant plasmid pNDV / rAI4-T4FHN-H9 is as follows figure 1 .
[0079] Construction step 1: Construction of a full-length expression clone expressing HA protein of avian influenza H9N2 subtype strain using chimeric Newcastle disease virus AI4-T4FHN as a vector
[0080] Biomaterial preparation:
[0081] A / chicken / jiangsu / WJ57 / 2012 (WJ57), the H9N2 subtype epidemic strain of avian influenza virus, was isolated and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University. The GenBank serial number is KJ000710.1
[0082] pCR2.1 vector: purchased from Invitrogen; Dephospharylation (BAP) Kit: purchased from Bao Biological Engineering (Dalian) Co., Ltd.; AMV reverse transcriptase, High fidelity DNA polymerase, T4 DNA ligase, AgaroseGel DNA Extraction Kit were purchased from Roche; Transfection reagent SuperFect and plasmid extraction kit (QIAprepSpin MiniPrep Kit) are product...
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