Chapter title |
PCR-Mediated Epitope Tagging of Genes in Yeast.
|
---|---|
Chapter number | 4 |
Book title |
Yeast Genetics
|
Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-1363-3_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1362-6, 978-1-4939-1363-3
|
Authors |
Radhika Mathur, Peter Kaiser, Mathur, Radhika, Kaiser, Peter |
Abstract |
Epitope tagging of genes is a powerful technique facilitating assays for gene function, determination of subcellular distribution of proteins, affinity purification, study of protein interaction with other proteins, DNA or RNA, and any other antibody-based approach in the absence of protein-specific antibodies. Here, we describe a one-step PCR-based strategy for insertion of epitope tags at the chromosomal locus. This method takes advantage of efficient homologous recombination in yeast. PCR amplified tags are directed to desired chromosomal loci with the help of primer-encoded flanking homologous sequences enabling selective epitope tagging of genes of interest. |
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