The Clining of the Whole Genome of CAV Isolate SJ1 from Shandong

The 1.5Kb fragment and the rest 0.8kb fragment of CAV isolate SJ1 from Shandong were amplified by two pairs of primers,CA5 CA6 and CA7 CA8,both of which contain EcoRI and BamHI sites respectively.The two fragments were cloned into EcoRI BamHI site of plasmid pQE32 and plasmid pBluescript(sk+),respectively.The 0.8kb fragment was then transrerred from pBluescript(sk+)to the former pQE32 by BamHI/KpnI digestion and subsequent ligation,to form plasmid as CAV pQE32 which contained the whole genome of CAV linearized at EcoRI sites.The CAV pQE32 was proved to be an infectious clone by transfection of MDCC MSB1 cell line The produced CAV virus detected both by immunofluence and ELISA.