Production of Mycobacterium tuberculosis MMAA4 protein in tobacco plant

Tuberculosis (TB), caused by Mycobacterium tuberculosis, is a vaccine-preventable disease still causing deaths of numerous humans. BCG, is the only licenced vaccine against TB. However, protection of this vaccine is insufficient among adults and elderlies. The studies on new and efficient anti-TB vaccines are ongoing. The aim of this study is to transfer the gene encoding MmaA4, a virulence factor of M. tuberculosis of which capacity for immune response stimulation was shown, and to provide the potential production of antigenic MmaA4 protein in tobacco plants for development of a recombinant TB vaccine. Fort his purpose, M. tuberculosis mmaA4 gene was cloned into pGFPGUS+ vector. Next, pGFPGUS+mmaA4 and pGFPGUS+ only as a control group were transferred into Agrobacterium tumefaciens. Tobacco plants were transformed via recombinant A. tumefaciens cells. The presence of mmaA4 gene was verified via PCR in transgenic tobacco plants transformed with GFPGUS+mmaA4. To show the production of recombinant MmaA4 protein in transgenic tobaccos, SDS-PAGE and Western blot analyses were performed. However, there was no difference between the transgenic plants carrying mmaA4 gene or not. Inability to observe the production of MmaA4 protein in transgenic tobaccos was attributed not to perform a codon optimization to express the bacterial gene in plant.