The Fluorescence Staining of Grass Carp Hemorrhage Virus(GCHV)Polypeptides

t had been reported that the GCHV(Gtass Carp Hemorrhage Virus)is a new member.in Reoviri-dae and has 11 polypeptides by our laboratory,We did further experiments on the viral polypeptides.The virions(GCHV-875)were purified from the medium of infection culture cells by CsCl density gradient centrifugation,dissolved in the buffer containing Tris-HCl 250mmol/L,glycerin 25%,SDS86mmol/L,2-ME 320mmol/L(pH6.9)and then heated at 100℃for 3 min.After that,by addingan equal volume of FITC solution(2mg/M)in 1 mol/L NaHCO3,the viral polypeptides were labelledwith FITC,and analysed by SDS-PAGE. All the viral structural polypeptides(11 bands),namelyVP130,VP120,VP113,VP107,VP75,VP65,VP60,VP52,VP42,VP39 and VP31,wete visible under. UV light.These polypeptides were retrieved by cutting down each band from the gel and demonstrated kept immunological property.The method can determine 500ng protein band in gel under UVlight.