CLONING AND EXPRESSION OF MCE1A GENE FROM BEIJING AND H37RV STRAIN FOR VACCINE CANDIDATE DEVELOPMENT.

BACKGROUND

Tuberculosis remains the leading cause of death in the world, especially wherever poverty, malnutrition and poor housing prevail. Beijing strain is the most common strain that causes tuberculosis in Indonesia. The wide spread of tuberculosis has been further aggravated by HIV-AIDS and drug resistance. Unfortunately, Bacille Calmette-Guerin (BCG) as the current vaccine has different protection function and efficacy. According to function analysis, gene was predicted to have a role in host invasion and survival of in human macrophages.

MATERIALS AND METHODS

We performed cloning and protein expression of gene of Beijing strain as local isolate and standard strain H37Rv as a comparison on the expression system BL21(DE3). gene from the strains were amplified by PCR and inserted into the vector pET28a. Each resulting recombinant plasmid was subsequently transformed into BL21(DE3) and Mce1A protein was expressed with IPTG induction.

RESULTS

BL21(DE3) was succesfully transformed with a recombinant plasmid that contains the gene insert with correct orientation and reading frame. There was no mutation found in the amino acids sequence for B and T cell epitope. Mce1A expression in BL21(DE3) showed a protein band that was higher than expected. The protein was confirmed with Western blotting using anti-His detector.

CONCLUSION

We assumed that Mce1A recombinant protein that has been expressed in BL21(DE3) is in their dimeric form or alternatively formed aggregates of different sizes.