Abstract
We have developed a high-throughput technology that allows parallel expression, purification, and analysis of large numbers of cloned cDNAs in the yeast Saccharomyces cerevisiae. The technology is based on a vector for intracellular protein expression under control of the inducible CUP1 promoter, where the gene products are fused to specific peptide sequences. These N-terminal and C-terminal epitope tags allow the immunological identification and purification of the gene products independent of the protein produced. By introducing the method of recombinational cloning we avoid time-consuming re-cloning steps and enable the easy switching between different expression vectors and host systems.
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© 2004 Humana Press Inc., Totowa, NJ
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Holz, C., Lang, C. (2004). High-Throughput Expression in Microplate Format in Saccharomyces cerevisiae . In: Balbás, P., Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 267. Humana Press. https://doi.org/10.1385/1-59259-774-2:267
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DOI: https://doi.org/10.1385/1-59259-774-2:267
Publisher Name: Humana Press
Print ISBN: 978-1-58829-262-9
Online ISBN: 978-1-59259-774-1
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