Cloning, expression, purification and preliminary oligomerisation analysis of recombinant protein Mycobacterium tuberculosis Rv1288

Aims: LysM containing-protein is widely distributed in all domains of life and this kind of protein is essential for various biological activities in living organisms. Rv1288, a LysM containing-protein with esterase, was found in Mycobacterium tuberculosis. Biophysical studies revealed that the protein is responsible for modulating lipid metabolism that enables pathogens to survive under extreme conditions and decrease the permeability of the pathogen's cell wall to drug therapeutic agents. However, recognition and interaction between the protein, lipid and carbohydrate moieties at the molecular level remains largely unknown and must be investigated. Therefore, a production of recombinant protein Rv1288 should be performed to aid the study. Methodology and results: In this study, we cloned the full-length cDNA of Rv1288 from M. tuberculosis strain H37Rv and expressed it in pET-24d-Escherichia coli BL21(DE3) cells. Affinity and size exclusion chromatography methods purified the protein, and its preliminary oligomerisation state was determined based on a calculated apparent molecular weight of the protein. Rv1288 was expressed as a soluble protein at 20 degrees C, induced with 1 mM of isopropyl beta-D-1-thiogalactopyranoside (IPTG). The calculated apparent molecular weight suggested that the Rv1288 protein formed a hexamer in solution. Conclusion, significance and impact of study: All the methods involved in this study to produce the recombinant Rv1288-pET24d and its soluble protein in E. coli cells have been described. Hence, it can be implemented for future studies.