Identification Of Nonessential Regions Of Genomic DNA Of Attenuated CPV Strain

Capripox.(Sheeppox and goatpox),casued by capripoxvirus,is an acute feverish and contagious disease in sheep or goat.Capripox is classified as list group A by Office International des Epizooties(OIE)and first group disease in China.Currently,studies on poxvirus vector vaccines are highlights.The genome of capri-pox virus has many features,including host specificity,a large number of nonessential Regions which are useful in inserting large exogenous genes.the capripox virus geno-me is 150kb in total length,In order to construct capripoxvirus vector vaccines,the primary tast identification of the nonessential regenions. The study was under take by random clonal method.The temperature-sensitive CPV strain was purfied by sucrose-density gradient.centrifugation then genomic DNA was extracted from The CPV virus. the fragements were randomly cloned into PUC18 and 6 postive clones PUA,PUB,PUC,PUD,PUE and PUF of Hindâ…¢d igested CPV DNA were obtained.Compared with the pellor(AY077835)strain, the nucleotide similarity was more than 90%.The P₁₁-P_7.5-LacZ report gene was inserted in the unique site of PUA,PUB,PUC,PUD,PUEand PUF,respectively.The plasmids PUA1,PUB1,PUC1,PUD1,PUE1,and PUF1 were acquired,which were transfected in BHK-21 infected by CPV.using blue–white plaque selection with existence of x-gal,one CPV fragment was identified as a nonessential region for CPV replication.