Analysis Of The Whole Genome Sequencing Of4NDV Predominant Strains And The Prokaryotic Expression Ducks NDV HN Protein

Newcastle disease (Newcastle Disease, ND) is an acute, highly contagious disease whichcaused by the Newcastle disease virus and capable of infecting a variety of birds, one of theimportant infectious diseases to harm the world poultry industry and resulting in hugeeconomic losses to many countries in the poultry industry.NDV has a variety of birdspathogenicity, except poultry, at least250species of birds can be natural or experimentalinfection of NDV. Genotype â…¦ NDVis of the most widely popular strains in our country.To research over genetic evolutionary trend of NDV in recent years, this articledetermination the full gene sequences of two genotype VII chicken Newcastledisease strains Chicken/China/SD04/2011and Chicken/China/SDWF07/2011, twogenotype â…¥ pigeon Newcastle disease strains Pigeon/China/SDLC/2011and Pigeon/China/SDS/2011by cloning and sequencing. The whole genome sequence andantigenic variation analysis showed that the nucleotide sequence homology are80%to87%between the four isolates and the china standard virulent F48E8, the foreign standardvirulent Herts/33and the vaccine strain chicken/N. Ireland/Ulster/67,B1, Clone30, LaSota, Mukteswar; the amino acid sequence is112RRQKRF117nearthe F protein precursor F0cleavage site and are in conformity with the velogen strainof NDV characteristics; two chicken source virus belongs to the â…¦d subtype mutant;twopigeon strain NP,P and L genes on certain amino acid changes may be relevant to their hostspecificity and virulence difference;antigenic variation analysis showed that negative reactionwith the monoclonal antibody1E5,initially speculated that not caused by HN protein of347amino acid variation,there may be other unknown factor determine the responsecharacteristics of the virus and monoclonal antibody1E5. The results ofPigeon/China/SDS/2011infection experimental to pigeon and chicken showed that it withstrong pathogenic to pigeon and weakly pathogenic to chicken.In2009, the laboratory isolated one Duck NDV SD03from the duck farms, sequenceanalysis and pathogenicity index measurement results show that it is a velogen strain. But the attack drug experiments to Cherry Valley duck showed that under the experimentalconditions SD03isolates do not cause disease to ducks, only detoxification. The HN proteinopen reading frame176~571amino acid (5receptor binding in the relevant area) is expressedby prokaryotic expression vector pET28a in the present experiment. It is a clear expressionband in the49kDa position that the expression product detected by SDS-PAGE. Expressionproduct detected by SDS-PAGE after purified by Ni-NTA affinity, then obtain pure targetrecombinant protein. The Western blotting analysis showed that the purified protein canspecific reaction with the monoclonal antibody of anti-NDV HN protein of antigenic sitesabout blood clotting properties and neutralizing activity and rabbit polyclonal, indicatingthat the recombinant protein has a good immunogenic. It lays the foundation of next furthermore preparation of monoclonal antibodies and in-depth study of its epitope preparations.