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Bacterial Expression System> | coli BL21(DE3)-pET28-kdpE (Table 1) and purified with Ni-NTA affinity column (GenScript).... Crude cell lysate was centrifuged to remove debris and supernatant was used to purify 6 × His KdpE by using affinity column (Ni-NTA, GenScript). | Get A Quote |
Kdp-ATPase is an inducible high affinity potassium uptake system that is widely distributed in bacteria, and is generally regulated by the KdpD/KdpE two-component system (TCS). In this study, conducted on Mycobacterium smegmatis, the kdpFABC (encoding Kdp-ATPase) expression was found to be affected by low concentration of K+, high concentrations of Na+, and/or [Formula: see text] of the medium. The KdpE was found to be a transcriptional regulator that bound to a specific 22-bp sequence in the promoter region of kdpFABC operon to positively regulate kdpFABC expression. The KdpE binding motif was highly conserved in the promoters of kdpFABC among the mycobacterial species. 5'-RACE data indicated a transcriptional... More