Abstract
We report here the discovery of 137 previously unappreciated genes in yeast through a widely applicable and highly scalable approach integrating methods of gene-trapping, microarray-based expression analysis, and genome-wide homology searching. Our approach is a multistep process in which expressed sequences are first trapped using a modified transposon that produces protein fusions to β-galactosidase (β-gal); non-annotated open reading frames (ORFs) translated as β-gal chimeras are selected as a candidate pool of potential genes. To verify expression of these sequences, labeled RNA is hybridized against a microarray of oligonucleotides designed to detect gene transcripts in a strand-specific manner. In complement to this experimental method, novel genes are also identified in silico by homology to previously annotated proteins. As these methods are capable of identifying both short ORFs and antisense ORFs, our approach provides an effective supplement to current gene-finding schemes. In total, the genes discovered using this approach constitute 2% of the yeast genome and represent a wealth of overlooked biology.
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Acknowledgements
We thank Lara Umansky, Stacy Piccirillo, and Sandra Matson for technical assistance, and Metin Bilgin for helpful suggestions. This work was supported by NIH Grant R01-CA77808 to M.S. A.K. is supported by a post-doctoral fellowship from the American Cancer Society.
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Kumar, A., Harrison, P., Cheung, KH. et al. An integrated approach for finding overlooked genes in yeast. Nat Biotechnol 20, 58–63 (2002). https://doi.org/10.1038/nbt0102-58
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DOI: https://doi.org/10.1038/nbt0102-58
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