Study On Molecular Biology Of Egg Drop Syndrome Virus HeBei Strain And Cloning Of The PVⅢ Gene

Egg Drop Syndrom Virus (EDSV) is the only member in the Fowl Adenovirus Group III, which has special biological characteristics. This test regardAV-127 strain,EDSV Yangzhou strain and EDSV Hebei strain as the basic material(candidate).Purify three duck embryo allantoic fluid of virus toget the comparatively pure virus sample.SDS-PAGE was run to analyse 3 virus samples after purifying on a column chroma-tography of Sephadex G-200, carry on virus albumen polypeptide analysis.The results show that the composition of polypetide was slightly different,and the number of polypepitides of EDSV Hebei strain, AV-127 and EDSV Yangzhou strain were 13,8,10 respectively.Restriction endonucleases(RE) maps and the size of fragments of three EDSV strains were compared and calculated by using six restriction endonucleases BamH I ,Hindâ…¢,EcoR I ,Pst I ,Sma I and Puv â…¡ .The results show that the RE maps of EDSV Hebei strain and AV-127 were similar for each other,and the maps of Sma â…  ,EcoR â…  Puvâ…¡ were same.The genomic DNA were co-digested with BamH I and other restriction enzymes.There are 6,6,9,7,5 fragments of EDSV Hebei strain gDNA digesting with BamH+Sma I, BamHâ… I + Pstâ…  ,BamH â…  + EcoRâ…  ,BamH â…  + Hindâ…¢ and BamHâ…  +Puvâ…¡.According to the nucleotide sequence of EDSV-AV-127 strain from the GeneBank ,a pair of oligonucleotide primer was designed by using Primer5.0.With use of polymerase chain reaction(PCR),the pâ…§ gene fragment was amplified from the genome of EDSV-Hebei strain and the target fragment directly was inserted into pBS-T vector ,the recombinant plasmid DNAs were used to transform the competent E. Coli JM109 cells.The positive colonies harboring the pâ…§ gene were indentified by digestion of restriction endonucleases and PCR. The positive colonies were sequenced.The result indicate that the plasimid contain the complete sequence of pâ…§ gene .Compared with AV-127 pâ…§ gene, there is no genetic variation. The results will facilitate the molecular virological study of EDSV and its engineering as an express vector.