ArticlePloS one2013
pAUL: a gateway-based vector system for adaptive expression and flexible tagging of proteins in Arabidopsis.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 25 citations in OpenAlex.
- Targeted translation inhibition of chloroplast and mitochondrial mRNAs by designer pentatricopeptide repeat proteins.Nucleic acids research · 2025Article
- Targeted introduction of premature stop codon in plant mitochondrial mRNA by a designer pentatricopeptide repeat protein with C-to-U editing function.The Plant journal : for cell and molecular biology · 2025Article
- SEC14-GOLD protein PATELLIN2 binds IRON-REGULATED TRANSPORTER1 linking root iron uptake to vitamin E.Plant physiology · 2023Article
- Shedding light on AT1G29480 ofPlant direct · 2022Article
- In vivo stabilization of endogenous chloroplast RNAs by customized artificial pentatricopeptide repeat proteins.Nucleic acids research · 2021Article
- Targeted misexpression of NAC052, acting in H3K4 demethylation, alters leaf morphological and anatomical traits in Arabidopsis thaliana.Journal of experimental botany · 2020Article
- Calcium-Promoted Interaction between the C2-Domain Protein EHB1 and Metal Transporter IRT1 Inhibits Arabidopsis Iron Acquisition.Plant physiology · 2019Article
- A versatile modular vector set for optimizing protein expression among bacterial, yeast, insect and mammalian hosts.PloS one · 2019Article
- Turnip Mosaic Virus Counteracts Selective Autophagy of the Viral Silencing Suppressor HCpro.Plant physiology · 2018Article
- The DnaJ-Like Zinc-Finger Protein HCF222 Is Required for Thylakoid Membrane Biogenesis in Plants.Plant physiology · 2017Article
- Selective autophagy limits cauliflower mosaic virus infection by NBR1-mediated targeting of viral capsid protein and particles.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- d-Lactate Dehydrogenase Links Methylglyoxal Degradation and Electron Transport through Cytochrome c.Plant physiology · 2016Article
- Production of dengue virus envelope protein domain III-based antigens in tobacco chloroplasts using inducible and constitutive expression systems.Plant molecular biology · 2016Article
- Two RNA recognition motif-containing proteins are plant mitochondrial editing factors.Nucleic acids research · 2015Article
- A zinc finger motif-containing protein is essential for chloroplast RNA editing.PLoS genetics · 2015Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Determination of protein function requires tools that allow its detection and/or purification. As generation of specific antibodies often is laborious and insufficient, protein tagging using epitopes that are recognized by commercially available antibodies and matrices appears more promising. Also, proper spatial and temporal expression of tagged proteins is required to prevent falsification of results. We developed a new series of binary Gateway cloning vectors named pAUL1-20 for C- and N-terminal in-frame fusion of proteins to four different tags: a single (i) HA epitope and (ii) Strep-tagIII, (iii) both epitopes combined to a double tag, and (iv) a triple tag consisting of the double tag extended by a Protein A tag possessing a 3C protease cleavage site. Expression can be driven by either the 35 S CaMV promoter or, for C-terminal fusions, promoters from genes encoding the chloroplast biogenesis factors HCF107, HCF136, or HCF173. Fusions of the four promoters to the GUS gene showed that endogenous promoter sequences are functional and drive expression more moderately and consistently throughout different transgenic lines when compared to the 35 S CaMV promoter. By testing complementation of mutations affected in chloroplast biogenesis factors HCF107 and HCF208, we found that the effect of different promoters and tags on protein function strongly depends on the protein itself. Single-step and tandem affinity purification of HCF208 via different tags confirmed the integrity of the cloned tags.
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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.