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US 6,207,407 B1Granted
Identification of antibacterial agents
AssigneeMillenium Pharmaceuticals, Inc.
InventorsWilliam Timberlake, Victoria Gavrais
Life of this patent
Estimating the term…
Abstract
A method for identifying a strain having a conditional lethal mutation which is essential for survival when strain is incubated under restrictive growth, conditions, and methods of identifying gene products and gene functions thereof.
Claims8
A method for identifying an antibacterial agent, the method comprising:
- (a) growing a mutagenized bacterial culture under first permissive conditions;
- (b) exposing the bacterial culture from step (a) to restrictive conditions for a period of time equivalent to at least two growth cycles;
- (c) exposing the bacterial culture from step (b) to second permissive conditions for a period of time equivalent to at least ten growth cycles;
- (d) selecting a mutated bacterial strain that survives step (a) but does not survive steps (b) and (c), thereby selecting a strain having a mutagenized gene carrying a conditional lethal mutation;
- (e) identifying a mutagenized gene product encoded by the mutagenized gene;
- (f) contacting a wild-type gene product target corresponding to the identified mutagenized gene product of step (e) with a test compound;
- and
- (g) detecting binding of the test compound to the wild-type gene product target as an indication that the test compound is an antibacterial agent.
5 more elements
A method for identifying an antibacterial agent, the method comprising:
- (a) growing a mutagenized bacterial culture under first permissive conditions;
- (b) exposing the bacterial culture from step (a) to restrictive conditions for a period of time equivalent to at least two growth cycles;
- (c) exposing the bacterial culture from step (b) to second permissive conditions for a period of time equivalent to at least ten growth cycles;
- (d) selecting a mutated bacterial strain that survives step (a) but does not survive steps (b) and (c), thereby selecting a strain having a mutagenized gene carrying a conditional lethal mutation;
- (e) identifying a mutagenized gene product encoded by the mutagenized gene;
- (f) contacting a wild-type gene product target corresponding to the identified mutagenized gene product of step (e) with a test compound;
- (g) detecting a decrease in activity of the wild-type gene product target in contact with the test compound, relative to activity of the wild-type gene product target in the absence of the test compound, as an indication that the test compound is an antibacterial agent.
4 more elements
A method for identifying an antibacterial agent, the method comprising:
- (a) growing a mutagenized bacterial culture under first permissive conditions;
- (b) exposing the bacterial culture from step (a) to restrictive conditions for a period of time equivalent to at least two growth cycles;
- (c) exposing the bacterial culture from step (b) to second permissive conditions for a period of time equivalent to at least ten growth cycles;
- (d) selecting a mutated bacterial strain that survives step (a) but does not survive steps (b) and (c), thereby selecting a strain having a mutagenized gene carrying a conditional lethal mutation;
- (e) identifying a mutagenized gene product encoded by the mutagenized gene;
- (f) contacting a wild-type gene product target corresponding to the mutagenized gene product identified in step (e) with a test compound;
- and
- (g) detecting a decrease in expression of the wild-type gene product target contacted with the test compound, relative to the level of expression in the absence of the test compound, as an indication that the test compound is an antibacterial agent.
5 more elements
Dependencies are inferred from the claim text.