Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 2452-2461 2452 Original Research Article https://doi.org/10.20546/ijcmas.2018.710.284 Cloning and Expression of Recombinant VP2 Capsid Protein Gene of Canine Parvovirus in E. coli System Sangeetha Subramani 1 , Hirak Kumar Mukhopadhyay 1 , Mouttou Vivek Srinivas 1* , Muthuraj Muthaiah 3 , Prabhakar Xavier Antony 1 and Jacob Thanislass 2 1 Department of Veterinary Microbiology, 2 Department of Veterinary Biochemistry, Rajiv Gandhi Institute of Veterinary Education & Research, Puducherry - 605 009, India 3 Microbiology Laboratory, TB and Chest Disease Hospital, Puducherry - 605 006, India *Corresponding author ABSTRACT Introduction Canine parvovirus (CPV) infection is one of the most important viral diseases in dogs. CPV infection is characterized by nausea, enteritis, leucopenia, and myocarditis in puppies (Appel et al., 1979). Canine parvovirus belongs to the genus Protoparvovirus, family Parvoviridae. The CPV virion is non-enveloped with icosahedral symmetry of 26nm diameter. It possesses a single stranded DNA genome of 5.2 kb in length. The virus has two open reading frame in its genome which encodes two non-structural (NS1 and NS2) and three structural (VP1, VP2 and VP3) proteins. VP1 contains the full length VP2 sequence plus an additional N-terminal domain. The VP2 capsid protein is a major protein and accounts for 90% of the viral capsid and is cleaved to VP3 by the host proteases. International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 7 Number 10 (2018) Journal homepage: http://www.ijcmas.com Canine parvovirus type - 2 (CPV-2) infection is one of the most important viral diseases in dogs and wild carnivores causing severe haemorrhagic gastroenteritis in young ones. VP2 capsid protein plays an important role in determining the antigenicity and diversity of the virus. Although, several CPV variants emerged but new CPV-2a is the predominant circulating field strain of CPV in India. In the present study, new CPV-2a field strain (KLD3) isolated in cell culture was selected and the whole CPV VP2 gene was used for the expression in the E. coli expression system. Prokaryote expressing monocistronic DNA cassette containing open reading frame of whole CPV capsid gene (VP2) downstream of T7 promoter was synthesized. Analysis of the expression in E. coli cells showed the presence of capsid protein. Recombinant capsid protein showed immunoreactivity similar to the whole CPV virus antigen, when reacted with polyclonal antibodies against the whole CPV virus particles. The use of indigenously developed recombinant protein, being very economical, can be used to develop field kit. As the recombinant protein is not infectious, use of it for CPV serodiagnostic assay is considered safe. Keywords Canine parvovirus type - 2 (CPV-2), E. coli, CPV Accepted: 18 September 2018 Available Online: 10 October 2018 Article Info