TY - JOUR
T1 - Towards establishing a method to screen for inhibitors of essential genes in mycobacteria
T2 - evaluation of the acetamidase promoter
AU - Raghunand, Tirumalai R.
AU - Bishai, William R.
AU - Chen, Ping
N1 - Funding Information:
We gratefully acknowledge the support of NIH grants AI 51668, AI36973, AI37856 and 3 AI43846. T.R.R. was the recipient of a Post-Doctoral fellowship from the Heiser Program.
PY - 2006/7
Y1 - 2006/7
N2 - As a successful pathogen, Mycobacterium tuberculosis has effectively infected one-third of the world's population. Despite the existence of compound libraries developed by recent advances in combinatorial chemistry, few compounds have been screened against M. tuberculosis. The use of a regulable promoter to control the level of expression of a drug target in living organisms has been shown to be advantageous compared with targetless whole-cell-based or in vitro biochemical screening approaches towards antibiotic discovery. In this study, we demonstrate that the acetamidase promoter from Mycobacterium smegmatis responds in a dose-dependent manner to different concentrations of its inducer acetamide. Using this promoter to regulate expression of a zeocin resistance gene in M. smegmatis, we show that the test strain exhibits increased sensitivity to zeocin at a low concentration of acetamide compared with a fully resistant phenotype at high doses of the inducer. This model system has indicated the feasibility of using a regulable promoter in designing a whole-cell-based high throughput screen for specific inhibitors against potential drug targets of M. tuberculosis.
AB - As a successful pathogen, Mycobacterium tuberculosis has effectively infected one-third of the world's population. Despite the existence of compound libraries developed by recent advances in combinatorial chemistry, few compounds have been screened against M. tuberculosis. The use of a regulable promoter to control the level of expression of a drug target in living organisms has been shown to be advantageous compared with targetless whole-cell-based or in vitro biochemical screening approaches towards antibiotic discovery. In this study, we demonstrate that the acetamidase promoter from Mycobacterium smegmatis responds in a dose-dependent manner to different concentrations of its inducer acetamide. Using this promoter to regulate expression of a zeocin resistance gene in M. smegmatis, we show that the test strain exhibits increased sensitivity to zeocin at a low concentration of acetamide compared with a fully resistant phenotype at high doses of the inducer. This model system has indicated the feasibility of using a regulable promoter in designing a whole-cell-based high throughput screen for specific inhibitors against potential drug targets of M. tuberculosis.
KW - Acetamidase promoter
KW - Dose-response
KW - Inhibitors
KW - Mycobacterium tuberculosis
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U2 - 10.1016/j.ijantimicag.2006.01.012
DO - 10.1016/j.ijantimicag.2006.01.012
M3 - Article
C2 - 16757153
AN - SCOPUS:33745186670
SN - 0924-8579
VL - 28
SP - 36
EP - 41
JO - International Journal of Antimicrobial Agents
JF - International Journal of Antimicrobial Agents
IS - 1
ER -