Correlation Engine 2.0
Clear Search sequence regions


  • HHT1 (1)
  • mCherry (1)
  • native (1)
  • protein domains (1)
  • protein tag (4)
  • RAD5 (1)
  • sfGFP (1)
  • target gene (2)
  • target proteins (1)
  • UBC13 (1)
  • yeast (2)
  • Sizes of these terms reflect their relevance to your search.

    Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique in the budding yeast Saccharomyces cerevisiae is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. However, several limitations hamper the application of this technique, such as the selectable markers not being reusable, tagging of only the C-terminal being possible, and a "scar" sequence being left in the genome. Here, we describe a strategy to solve these problems by tagging target genes based on a pop-in/pop-out and counter-selection system. Three fluorescent protein tag (mCherry, sfGFP, and mKikGR) and two epitope tag (HA and 3×FLAG) constructs were developed and utilized to tag HHT1, UBC13 or RAD5 at the chromosomal locus as proof-of-concept.

    Citation

    Qian Wang, Huijun Xue, Siqi Li, Ying Chen, Xuelei Tian, Xin Xu, Wei Xiao, Yu Vincent Fu. A method for labeling proteins with tags at the native genomic loci in budding yeast. PloS one. 2017;12(5):e0176184

    Expand section icon Mesh Tags

    Expand section icon Substances


    PMID: 28459859

    View Full Text