Molecular Cloning and Restriction Endonuclease Analysis of Canine Parvovirus DNA Amplified by Polymerase Chain Reaction

Canine parvovirus 2 (CPV-2) is a single stranded DNA virus of Parvoviridae family that causes enteritis and myocarditis in canines. DNA was isolated from an isolate of CPV-2 and 765 bp sequence of VP-2 gene was amplified by PCR using VP2 specific primer. The amplification was highly efficient as evident by single DNA band in agarose gel. The PCR product generated was ligated in pGEM-T Easy vector by TA cloning. The recombinant plasmid DNA was isolated and digested with Bam H I and Sal I restriction enzymes, which yielded the fragment of vector DNA of 3015 bp and the insert of 765 bp.The insert was further characterized by restriction enzyme (RE) mapping using Hha I. It was concluded that PCR along with RE analysis of amplicons can be employed successfully for detection and characterization of CPV which in turn would help in proper and effective management and control of the disease.