Vet World Vol.14 June-2021 Article-35
Research Article
Veterinary World, 14(6): 1682-1688
https://doi.org/10.14202/vetworld.2021.1682-1688
Expression of recombinant 35 kDa fragment of VP2 protein of canine parvovirus using Escherichia coli expression system
2. Center of Excellence on Agricultural Biotechnology: (AG-BIO/PERDO-CHE), Bangkok 10900, Thailand.
3. Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok 10900, Thailand.
4. Molecular Diagnostic Laboratory, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
5. Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
6. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
Background and Aim: Canine parvovirus (CPV) is one of the most common viral infections in dogs, causing acute hemorrhagic gastroenteritis and high mortality. Vaccination effectively prevents CPV infection. However, the currently available CPV vaccines have concerns such as maternal immunity interference, shedding of virus vaccine, and false-positive result based on polymerase chain reaction after vaccination. A subunit vaccine can overcome these problems. This study aimed to express the recombinant 35 kDa fragment of the VP2 protein (consisting of epitopes 1-7) and the recombinant full-length VP2 protein (consisting of epitopes 1-10) and to study the ability of these two recombinant proteins to react with rabbit anti-CPV polyclonal antibodies.
Materials and Methods: The full length and 35 kDa fragment of VP2 gene of CPV were cloned into the pBAD202 Directional TOPOTM expression vector and expressed in E. coli. The recombinant full-length and the recombinant 35 kDa fragment proteins of VP2 were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting.
Results: The recombinant full-length and the recombinant 35 kDa fragment VP2 genes were successfully cloned and expressed. The optimum concentrations of arabinose and induction time for the recombinant full-length and the recombinant 35 kDa fragment VP2 proteins were 0.2% for 6 h and 0.02% for 6 h, respectively. The recombinant full-length and the recombinant 35 kDa fragment VP2 protein molecular weights were approximately 81 and 51 kDa, respectively. The recombinant full-length and the recombinant 35 kDa fragment VP2 proteins specifically interacted with rabbit anti-CPV polyclonal antibodies.
Conclusion: These results suggest that the recombinant 35 kDa fragment and the recombinant full-length VP2 proteins may be useful in developing a CPV diagnostic test or vaccine. Keywords: canine parvoviruses, Escherichia coli expression system, recombinant protein, VP2 gene.
Keywords: canine parvoviruses, Escherichia coli expression system, recombinant protein, VP2 gene.
How to cite this article: Inthong N, Kaewmongkol S, Meekhanon N, Suwan E, Sricharern W, Satchasataporn K, Sinsiri R, Sirinarumitr K, Sirinarumitr T (2021) Expression of recombinant 35 kDa fragment of VP2 protein of canine parvovirus using Escherichia coli expression system, Veterinary World, 14(6): 1682-1688.
Received: 18-01-2021 Accepted: 11-05-2021 Published online: 29-06-2021
Corresponding author: Theerapol Sirinarumitr E-mail: fvettps@yahoo.com
DOI: 10.14202/vetworld.2021.1682-1688
Copyright: Inthong, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.