Anuj Kumar
Project Manager, Yale Genome Analysis Center
Department of Molecular, Cellular and Developmental Biology
Yale University
P. O. Box 208103
New Haven, CT 06520-8103
telephone: 203-429-9949
fax: 203-432-6161
email: anuj.kumar@yale.edu
prestype: Platform
presenter: Anuj Kumar
Transposable elements are powerful tools for insertional mutagenesis, notable for the scale of throughput and ease of analysis they offer. As a result, transposon-based systems provide an excellent method of genome-wide mutagenesis. Illustrating this point, we have employed transposon mutagenesis as a means of elucidating gene function on a genome-wide scale in Saccharomyces cerevisiae. Our approach is unique in its versatility: by designing specially modified multi-functional mini-transposons, we can measure gene expression, generate gene disruptions, and localize gene products; all from a single transposon insertion event. Using these multi-functional transposons, we have generated a collection of roughly 17,000 yeast mutants, each carrying a single transposon insertion within a region of the genome expressed during vegetative growth and/or sporulation. This collection has been used to determine disruption phenotypes for nearly 8,000 strains under 20 different growth conditions. Additionally, we have identified over 300 previously non-annotated open reading frames and determined the subcellular localization of transposon-tagged proteins in approximately 9,000 yeast mutants. In total, our study encompasses over a quarter of a million data points, providing insight into the function of nearly 3,000 yeast genes. In addition to the wealth of data generated by this approach, our genome-wide collections of defined mutants constitute a valuable laboratory reagent, streamlining future genetic screens by eliminating the need to identify the affected gene within a given mutant of interest. As such, these collections of defined alleles promise to greatly expedite studies of gene function on a genomic scale.
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