Expression and epitope prediction of MPT64 recombinant proteins from clinical isolates of as immunoserodiagnostic candidates.

BACKGROUND AND AIM

The success in the handling and prevention of tuberculosis (TB) cases is highly dependent on their rapid detection, monitoring, and treatment. The efficacy of the Bacille Calmette-Guerin (BCG) vaccine is inconclusive in eastern Indonesia. The gene of encodes an antigenic protein that is considered to be a virulence factor, as it can stimulate the immune response in patients with TB. This study aimed to study the expression and epitope indicator of MPT64 recombinant proteins from clinical isolates of as immunoserodiagnostic candidates for pET SUMO plasmids from clinical isolates as candidates for serodiagnostic tests and recombinant vaccines.

MATERIALS AND METHODS

The polymerase chain reaction (PCR) product of the gene was inserted into the SUMO pET plasmid, which was then transformed into BL21 (DE3) cells and expressed in Luria Bertani media induced by 1.0 M IPTG. Subsequently, sequencing was performed and the results were analyzed using the ClustalW and National Center for Biotechnology Information BLAST software. The T-cell epitope prognosis was then explained by GENETYX version 8.0., for the prediction of B-cell epitope, as assessed using an Immune Epitope Database analysis.

RESULTS

The PCR product of the gene had a length of 619 bp. Moreover, SDS- polyacrylamide gel electrophoresis and Western blotting revealed that the protein encoded by the gene weighed 36 kDa. We gained nine specific T-cell epitopes according to Iad Pattern position and eight epitopes according to Rothbard/Taylor Pattern Position; furthermore, we detected five B-cell epitopes in the gene.

CONCLUSION

The MPT64 protein encoded by the gene carries epitopes that are realized by lymphocytes and represent potential immunoserodiagnostic candidates in diagnostic immunology.