The Genome Sequencing, Full-length CDNA Cloning Of Rabbit Hemorrhagic Disease Virus WHNRH Strain And The Construction Of Expression Vectors Of VP60 And Full-length CDNA Of RHDV

Rabbit hemorrhagic disease was an acute and fatal contagious disease for rabbits caused by rabbit hemorrhagic disease virus(RHDV) and caused heavy economy losses.The disease first broke out in China in 1984 and until now it existed all over China. The molecular epidemiology of RHD in China was in deficiency and there only one genome sequence of RHDV in GenBank from China until 2004. The sequencing progress had not been reported in details before in China. The reverse genetics and DNA vaccine research had not beeen reported in the world.So the research included the following contents:1.Isolation and identification of RHDV WHNRH strain: Liver tissues from animals that were suspected to have died of rabbit hemorrhagic disease (RHD) were used for isolation and identification of the causative agent. Since there is no cell culture system to grow RHDV, animals were used for virus isolation in the study. The suspected causative agent was inoculated and went through rabbits three times,then the inoculated rabbits died regularly and the pathological changes were identical to RHD after third passage in rabbitsA method purifying RHDV with chloroform and polyethylene glycol 6000 was applied to treat the liver tissues of died rabbits. The causative agent isolated as a preparation reacted positively by hemagglutination (HA) assays with 216 hemagglutination units and the hemagglutination was inhibited by serum against RHDV specially.In agar diffusion there form clear precipitate lines between the preparation and positive serum against RHDV.The preparation was negative-stained with phosphotungstic acid and then observed with electron microscope, many virion without peplos measured approximately 30-40 nm in diameter was obtained. Morphologically, the isolated virus were identical to RHDV.A pair of...