[Drug screening model acting on out-membrane protein OprM in pseudomonas aeruginosa efflux pump system].
OBJECTIVE
To establish an efflux pump inhibitor screening model with the out-membrane protein OprM in Pseudomonas aeruginosa efflux pump system as the target point.


METHODS
Efflux pump out-membrane protein gene oprM was obtained from standard Pseudomonas aeruginosa PA01 strain. Expression of OprM protein was induced in E. coli strain HS151 with T-easy vector as the cloning vector, and pMMB67EH as the expression vector. In order to evaluate the function of OprM protein, we measured intracellular tetracycline concentrations with liquid scintillation counter, measured the diameters of bacteriostatic circles with paper disc, and then established a screening model accordingly.


RESULTS
OprM protein was highly expressed. Using Pseudomonas aeruginosa as the main detecting bacteria, we established a drug screening model acting on OprM. A total of 1 600 microbial fermentation samples were screened with this model, among which 56 positive strains were found, with a positive rate of 3.5%.


CONCLUSIONS
OprM plays an important role in drug efflux. The established model has good specificity and maneuverability.
Tian
Rui
R
National Laboratory for Screening New Microbial Drugs, Institute of Medicinal Biotechnology, CAMS and PUMC, Beijing 100050, China.
Yu
Li-yan
LY
Xiao
Chun-ling
CL
Zuo
Lian
L
Yao
Tian-jue
TJ
Yang
Li-xia
LX
D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004352:Drug Resistance, Microbial; D018432:Drug Resistance, Multiple / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D026901:Membrane Transport Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics*
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; C402263:OprM protein, Pseudomonas aeruginosa