Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2003-07-11
2008-08-19
Kim, Young J. (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C536S023100, C435S183000
Reexamination Certificate
active
07413856
ABSTRACT:
A method and assay system is provided for screening for antimicrobial agents that inhibit or otherwise disrupt the transcription of bacterial aminoacyl-tRNA synthetases, particularly those in Gram-positive bacteria. The method and assay utilizes purified RNA polymerase, isolated, for example, from eitherBacillus subtilisorEscherichia coli, and a template DNA fragment which encodes glycyl-tRNA synthetase, containing the promoter and leader region of theB. subtilisglyQS gene, including the T-Box terminator/antiterminator. Incubation in the presence of nucleotide triphosphates results in synthesis of an RNA transcript initiating at the glyQS promoter and terminating at the leader region terminator. Read-through of the terminator, and synthesis of an extended transcript, is dependent on addition of purified glycyl-tRNA. The method and assay is dependent on the use of the specified tRNA species; non-specific tRNA fails to stimulate antitermination. The assay can be used to screen for inhibitors that specifically inhibit the T-box antitermination mechanism.
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Grundy Frank J.
Henkin Tina M.
Calfee Halter & Griswold LLP
Kim Young J.
The Ohio State University Research Foundation
Woolwine Samuel
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