Other

Basf Aktiengesellschaft

American Cyanamid Company

Sanford Jay Silverman

Donald R Kirsch

David Botstein

Margaret Hsien-Fen Kuh Lai

Robert Scott McIsaac

Marcus Noyes

A system allows for rapid and specific induction of individual genes in eukaryotic cells using a chimeric transcriptional activator that is responsive to hormone inducer. Upon addition of the hormone, cytoplasmic transcriptional activator localizes to the nucleus and subsequently binds to promoters containing sequences that bind to its DNA-binding domain. Genetic modifications allow for rapid and specific degradation of a targeted protein upon addition of hormone by means of a regulated degron method that utilizes a protease variant. This system is useful for discovering new compounds by high throughput screening when introducing compound libraries to these protein-depleted cells.

Systems for induction of gene expression and protein depletion in yeast

A method for the identification of agents which inhibit chitin synthesis, thus exhibiting potential fungicidal and insecticidal activity, involves the incubation of test samples in neutral cultures containing calcofluor white, a fluorochrome that causes lethal chitin hyper-polymerization in the yeast. Cultures containing samples that inhibit chitin synthesis exhibit enhanced growth because the growing fungus is rescued from the adverse effects of calcofluor white. In the practice of the invention, the test sample is added to a culture or culture area containing calcofluor white and the culture is incubated with the test sample for such time under such conditions sufficient to observe yeast cell growth inhibition in a corresponding culture or culture area containing calcofluor but no test sample. The extent of growth in the culture or culture area containing test sample is then compared with the extent of growth in the culture or culture area containing no test sample, and the presence of chitin synthesis inhibition is determined by observation of whether culture growth in the presence of test sample exceeds growth in its absence. Preferred embodiments employ test samples on disks or in wells in solidified cultures and a known chitin synthesis inhibitor as a positive control which is compared to the test sample.

Calcafluor screen for chitin biosynthesis inhibitors

The present invention is directed to a method for screening samples for the identification of agents exhibiting potential fungicidal and insecticidal activity for a wide variety of agricultural, medical and pharmaceutical uses. The method utilizes cells that comprise a plasmid-born CTS gene of Saccharomyces cerevisiae, which allows for over expression of chitinase. Compounds that inhibit a hydrolyic action on methyl-umbelliferyltriacetyl chitotriose, but which are not toxic to the cells are detected by decrease in conversion of substrate

Screen for inhibitors of chitinase

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Roosevelt, NJ, United States of America

Sanford Jay Silverman