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Additional modulesfor versatile andeconomical PCR-basedgene deletion andmodification inSaccharomycescerevisiae Mark S. LongtineAmos MckenzieDouglas J. DeMarini1998 год

Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae
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Аннотация: An important recent advance in the functional analysis of Saccharomyces cerevisiae genes is the development of the one-step PCR-mediated technique for deletion and modification of chromosomal genes. This method allows very rapid gene manipulations without requiring plasmid clones of the gene of interest. We describe here a new set of plasmids that serve as templates for the PCR synthesis of fragments that allow a variety of gene modifications. Using as selectable marker the S. cerevisiae TRP1 gene or modules containing the heterologous Schizosaccharomyces pombe his5+ or Escherichia coli kanr gene, these plasmids allow gene deletion, gene overexpression (using the regulatable GAL1 promoter), C- or N-terminal protein tagging [with GFP(S65T), GST, or the 3HA or 13Myc epitope], and partial N- or C-terminal deletions (with or without concomitant protein tagging). Because of the modular nature of the plasmids, they allow efficient and economical use of a small number of PCR primers for a wide variety of gene manipulations. Thus, these plasmids should further facilitate the rapid analysis of gene function in S. cerevisiae. © 1998 John Wiley & Sons, Ltd.
Год издания: 1998
Авторы: Mark S. Longtine, Amos Mckenzie, Douglas J. DeMarini, Nirav G. Shah, Achim Wach, Arndt Brachat, Peter Philippsen, John R. Pringle
Издательство: Wiley
Источник: Yeast
Ключевые слова: Fungal and yeast genetics research, RNA and protein synthesis mechanisms, Bacterial Genetics and Biotechnology
Другие ссылки: Yeast (HTML)
PubMed (HTML)