Cloning and expression of P67 protein of Mycoplasma leachii
| Autor: | Bhavani Puvvala, Rajneesh Rana, Valsala Rekha, Sabarinath Thankappan, Arun Thachappully Remesh, Viswas Konasagara Nagaleekar |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Veterinary medicine 030106 microbiology cloning Dot blot SF1-1100 law.invention P67 protein 03 medical and health sciences Western blot Affinity chromatography law Mycoplasma leachii expression SF600-1100 medicine Polyacrylamide gel electrophoresis Polymerase chain reaction western blot General Veterinary medicine.diagnostic_test Chemistry Fusion protein Molecular biology Animal culture genomic DNA dot blot Recombinant DNA Research Article |
| Zdroj: | Veterinary World, Vol 10, Iss 9, Pp 1108-1113 (2017) Veterinary World |
| ISSN: | 2231-0916 0972-8988 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|