A protocol for cloning, expression, and purification of Lysine exporter (LysE) gene of Mycobacterium tuberculosis

Background: Tuberculosis (TB) is among the deadliest diseases and a significant cause of illness across globe. Several studies on mycobacterial proteins, such as proteases transporters that are essential for survival pathogenesis have aimed to develop an efficient anti-tubercular agent. In mycobacterium, lysine exporter (LysE) amino acid transporter probable target agent it responsible bacterial growth inhibition also absent in widely used Bacillus Calmette-Guérin (BCG) vaccine. Methods: Some purified LysE using different protocols. This study describes protocol purifying constructs LysE, focusing its hydrophobic region immobilized metal affinity chromatography (IMAC) after expressing gene expression system. pET vector (pET28a) vector. Amplied ligated with pET28a vector, resultant plasmid then transformed into E. coli cells. The has histidine tag makes purification process convenient. After IMAC, samples will be subjected size-exclusion further purification. Results: Cloning amplification findings analyzed 1% agarose gel, protein outcomes examined sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Domain-specific can agent. Conclusions: Despite being potential target, research quite limited this protein. Therefore, we aim purify analysis. Similar protocols already been implemented several other proteins >95% purity.