ArticleGenetics2015
Avoiding the ends: internal epitope tagging of proteins using transposon Tn7.
Article in Genetics, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- EpicTope: predicting and validating non-disruptive epitope tagging sites.Development (Cambridge, England) · 2026Article
- Detection and Localization of Type II Membrane Proteins by Endogenous Gene Tagging and Confocal Microscopy.Methods in molecular biology (Clifton, N.J.) · 2026Article
- LncRNA-encoded peptides in cancer.Journal of hematology & oncology · 2024Review
- Solid-Phase Cell-Free Protein Synthesis and Its Applications in Biotechnology.Advances in biochemical engineering/biotechnology · 2023Article
- The Emerging Role of uORF-Encoded uPeptides and HLA uLigands in Cellular and Tumor Biology.Cancers · 2022Review
- Small open reading frames in plant research: from prediction to functional characterization.3 Biotech · 2022Review
- When Long Noncoding Becomes Protein Coding.Molecular and cellular biology · 2020Review
- Membrane phospholipid alteration causes chronic ER stress through early degradation of homeostatic ER-resident proteins.Scientific reports · 2019Article
- Article
- Global analysis of protein homomerization inGenome research · 2019Article
- Universal intracellular biomolecule delivery with precise dosage control.Science advances · 2018Article
- Sequence-based prediction of permissive stretches for internal protein tagging and knockdown.BMC biology · 2017Article
- Subcellular Localization of Pseudomonas syringae pv. tomato Effector Proteins in Plants.Methods in molecular biology (Clifton, N.J.) · 2017Article
- Scarless Gene Tagging with One-Step Transformation and Two-Step Selection in Saccharomyces cerevisiae and Schizosaccharomyces pombe.PloS one · 2016Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Peptide tags fused to proteins are used in a variety of applications, including as affinity tags for purification, epitope tags for immunodetection, or fluorescent protein tags for visualization. However, the peptide tags can disrupt the target protein function. When function is disrupted by fusing a peptide to either the N or C terminus of the protein of interest, identifying alternative ways to create functional tagged fusion proteins can be difficult. Here, we describe a method to introduce protein tags internal to the coding sequence of a target protein. The method employs in vitro Tn7-transposon mutagenesis of plasmids for random introduction of the tag, followed by subsequent Gateway cloning steps to isolate alleles with mutations in the coding sequence of the target gene. The Tn7-epitope cassette is designed such that essentially all of the transposon is removed through restriction enzyme digestion, leaving only the protein tag at diverse sites internal to the ORF. We describe the use of this system to generate a panel of internally epitope-tagged versions of the Saccharomyces cerevisiae GPI-linked membrane protein Dcw1 and the Candida glabrata transcriptional regulator Sir3. This internal protein tagging system is, in principle, adaptable to tag proteins in any organism for which Gateway-adapted expression vectors exist.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.