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Yeast Consortium

High-throughput flow-cytometric multiplexed yeast assays for discovery, pathway analysis, and chemical screening.

UNMCMD is actively seeking collaborations and assay providers for new chemical screens against defined biological targets as part of the NIH Roadmap Initiative. UNMCMD offers access to the NIH Small Molecule Repository (300K+ small molecules) and HTS capabilities.

Consortium members (partial list):

Poster: (presented at the 2008 Yeast Genetics and Molecular Biology Meeting.)

High-throughput flow-cytometric multiplexed yeast assays for discovery, pathway analysis, and chemical screening.

[See full poster PDF...]

Chris Allen1, Bruce Edwards1, Susan Young1, Peter Simons1, Mark Carter1, Anna Waller1, Kelly Trujillo6, MaryAnn Osley6, Ray Joe2, Mabel Padilla2, Phillip Tapia2, Sushmita Roy3, Maggie Werner-Washburne2, Claudio De Virgilio7, Hong Cai4, Larry Sklar1,5. 1) Cytometry and Cancer Center, U. New Mexico School of Medicine; 2) Biology Department, U. New Mexico; 3) Computer Science Department, U. New Mexico, Albuquerque, NM 87131; 4) National Flow Cytometry Resource, Los Alamos, NM 87545; 5) Dept. of Pathology, 6) Dept. of Molecular Genetics and Microbiology, U. New Mexico School of Medicine, Albuquerque, NM 87131, 7) Department of Medicine, Department of Biochemistry, University of Fribourg, Fribourg, Switzerland.

Abstract: The flow cytometry analytical platform is well-suited for multiplexing applications, enabling simultaneous quantitative analysis in individual cells or particles labeled with optical markers of biochemical expression or physiological response. Until recently, throughput in conventional flow cytometry has been limited by sampling time and analytical speed. Improvements in automation technology (384 wells sampled in ~10 minutes) and adaptation of assay formats for multiplexing targets (tens at a time) and/or multiplexing compounds (thousands at a time) have enabled the use of flow cytometry as a discovery tool for high content/ high throughput screening of large compound libraries. We are combining extant and emerging yeast technologies with our expertise in HT flow cytometry to create sophisticated platforms for HTS. For example, the yeast twohybrid system has recently been adapted and optimized for flow cytometric analysis. Both the speed of detection for protein-protein interactions and enrichment of positive interactors by magnetic sorting have been greatly enhanced. Second, we completed an initial screen of the yeast GFP collection under exponential and carbon-starvation growth conditions and identified several proteins that were highly expressed in glucose-exhausted cultures. A third example is an assay for DNA-protein biomolecular interactions in which overexpressed GST-fusion proteins are purified and bound to GSH-conjugated flow-cytometry beads in multiplex format.

To inquire regarding collaboration and consortium membership, contact: Christopher Allen.

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