ArticlePlant methods2013
Optimizing plant transporter expression in Xenopus oocytes.
Article in Plant methods, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 22 citations in OpenAlex.
- The rice transcription factor Nhd1 regulates root growth and nitrogen uptake by activating nitrogen transporters.Plant physiology · 2022Article
- Structures and General Transport Mechanisms by the Major Facilitator Superfamily (MFS).Chemical reviews · 2021Article
- Harnessing a Transient Gene Expression System inPlants (Basel, Switzerland) · 2021Article
- Overexpression of the High-Affinity Nitrate TransporterInternational journal of molecular sciences · 2020Article
- An Iron-Activated Citrate Transporter, MtMATE67, Is Required for Symbiotic Nitrogen Fixation.Plant physiology · 2018Article
- NRT1.5/NPF7.3 Functions as a Proton-Coupled HThe Plant cell · 2017Article
- Introducing an algal carbon-concentrating mechanism into higher plants: location and incorporation of key components.Plant biotechnology journal · 2016Article
- Plant Aquaporins: Genome-Wide Identification, Transcriptomics, Proteomics, and Advanced Analytical Tools.Frontiers in plant science · 2016Review
- Knock-Down of a Tonoplast Localized Low-Affinity Nitrate TransporterFrontiers in plant science · 2016Article
- Identification and functional assay of the interaction motifs in the partner protein OsNAR2.1 of the two-component system for high-affinity nitrate transport.The New phytologist · 2014Article
- Chrysanthemum CmNAR2 interacts with CmNRT2 in the control of nitrate uptake.Scientific reports · 2014Article
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Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
Abstract
backgroundRapid improvements in DNA synthesis technology are revolutionizing gene cloning and the characterization of their encoded proteins. Xenopus laevis oocytes are a commonly used heterologous system for the expression and functional characterization of membrane proteins. For many plant proteins, particularly transporters, low levels of expression can limit functional activity in these cells making it difficult to characterize the protein. Improvements in synthetic DNA technology now make it quick, easy and relatively cheap to optimize the codon usage of plant cDNAs for Xenopus. We have tested if this optimization process can improve the functional activity of a two-component plant nitrate transporter assayed in oocytes.
resultsWe used the generally available software (http://www.kazusa.or.jp/codon/; http://genomes.urv.es/OPTIMIZER/) to predict a DNA sequence for the plant gene that is better suited for Xenopus laevis. Rice OsNAR2.1 and OsNRT2.3a DNA optimized sequences were commercially synthesized for Xenopus expression. The template DNA was used to synthesize cRNA using a commercially available kit. Oocytes were injected with cRNA mixture of optimized and original OsNAR2.1 and OsNRT2.3a. Oocytes injected with cRNA obtained from using the optimized DNA template could accumulate significantly more NO3- than the original genes after 16 h incubation in 0.5 mM Na15NO3. Two-electrode voltage clamp analysis of the oocytes confirmed that the codon optimized template resulted in significantly larger currents when compared with the original rice cDNA.
conclusionThe functional activity of a rice high affinity nitrate transporter in oocytes was improved by DNA codon optimization of the genes. This methodology offers the prospect for improved expression and better subsequent functional characterization of plant proteins in the Xenopus oocyte system.
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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.