The Ohio State University

Ohio University

Ahio University

Frank J Grundy

Tina M Henkin

Stephen Bergmeier

Jennifer V Hines

Compounds that are selective binding agents specific for RNA secondary and tertiary structures and a fluorescence assay for detecting the binding of small molecules to T-box antiterminator model mRNAs. The compounds include novel tri-substituted oxazolidinones and amino alcohols. The assay provides fluorescently labeled model mRNAs that include conserved sequence information typical of antiterminator bulge regions found in the mRNA of T-box gene regulatory systems.

Compositions that bind antiterminator RNA and assay for screening for such compositions

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 eitheror, and a template DNA fragment which encodes glycyl-tRNA synthetase, containing the promoter and leader region of theglyQS 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.

In vitro transcription assay for T box antitermination system

Compositions that bind antiterminator RNA and assay for screening such compositions

Growing community of inventors

Dublin, OH, United States of America

Frank J Grundy