Cloning and expression of P67 protein of Mycoplasma leachii .
- Resource Type
- Academic Journal
- Authors
- Thankappan S; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.; Rana R; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.; Remesh AT; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.; Rekha V; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.; Nagaleekar VK; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.; Puvvala B; Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.
- Source
- Publisher: Veterinary World Country of Publication: India NLM ID: 101504872 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0972-8988 (Print) Linking ISSN: 09728988 NLM ISO Abbreviation: Vet World Subsets: PubMed not MEDLINE
- Subject
- Language
- English
- ISSN
- 0972-8988
Aim: The present study was undertaken to clone, express and study the immunogenicity of P67 protein of Mycoplasma leachii .
Materials and Methods: P67 gene was amplified from genomic DNA of M. leachii . The polymerase chain reaction (PCR) product was inserted in pRham N-His SUMO Kan vector and was used to transform competent Escherichia cloni 10G cells. Recombinant protein expression was done by inducing cells with 0.2% Rhamnose. Purification was done using nickel nitrilotriacetic acid affinity chromatography. Western blot and dot blot analysis were performed to assess the immunoreactivity of P67 protein.
Results: PCR amplicon size of P67 gene was found to be 1500 base pair. The size of the fusion protein with SUMO tag was 79 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The recombinant P67 fusion protein expressed in pRham N-His SUMO Kan vector was found to be immunogenic in both western blot and dot blot analysis.
Conclusion: Western blot and dot blot analysis of P67 protein of M. leachii revealed that the protein is immunogenic. Further work is needed to evaluate the role of P67 antigen of M. leachii as an immunodiagnostic agent.