ArticlePloS one2012
Robust expression and secretion of Xylanase1 in Chlamydomonas reinhardtii by fusion to a selection gene and processing with the FMDV 2A peptide.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 81 papers.
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Who cites it
81 citing papers in PubMed, 241 citations in OpenAlex.
- An F2A sequence permits correct localization of a secreted and a nuclear localized reporter inmicroPublication biology · 2026Article
- Expression and Characterization of Recombinant Triple Laterosporulin in Chlamydomonas reinhardtii.Probiotics and antimicrobial proteins · 2026Article
- A game of tag: A review of protein tags for the successful detection, purification and fluorescence labelling of proteins expressed in microalgae.The Plant journal : for cell and molecular biology · 2025Review
- Engineering the green algae Chlamydomonas incerta for recombinant protein production.PloS one · 2025Article
- Efficient secretory production of recombinant proteins in microalgae using an exogenous signal peptide.Frontiers in microbiology · 2025Article
- Harnessing the potential of microalgae for the production of monoclonal antibodies and other recombinant proteins.Protoplasma · 2024Review
- Establishing the green algaebioRxiv : the preprint server for biology · 2024Article
- Proteomic analysis reveals molecular changes following genetic engineering in Chlamydomonas reinhardtii.BMC microbiology · 2024Article
- Bioinformatic Prediction and High Throughput In Vivo Screening to Identify Cis-Regulatory Elements for the Development of Algal Synthetic Promoters.ACS synthetic biology · 2024Article
- Type II metacaspase mediates light-dependent programmed cell death in Chlamydomonas reinhardtii.Plant physiology · 2024Article
- Comparing the Ability of Secretory Signal Peptides for Heterologous Expression of Anti-Lipopolysaccharide Factor 3 inMarine drugs · 2023Article
- Secretory Expression and Application of Antilipopolysaccharide Factor 3 inBioengineering (Basel, Switzerland) · 2023Article
- Site-specific gene knock-in and bacterial phytase gene expression inFrontiers in plant science · 2023Article
- Microalgae as an Efficient Vehicle for the Production and Targeted Delivery of Therapeutic Glycoproteins against SARS-CoV-2 Variants.Marine drugs · 2022Review
- Increased Lipids inInternational journal of molecular sciences · 2022Article
- Towards green biomanufacturing of high-value recombinant proteins using promising cell factory: Chlamydomonas reinhardtii chloroplast.Bioresources and bioprocessing · 2022Review
- Unassembled cell wall proteins form aggregates in the extracellular space of Chlamydomonas reinhardtii strain UVM4.Applied microbiology and biotechnology · 2022Article
- Structural Elements of DNA and RNA Eukaryotic Expression Vectors for In Vitro and In Vivo Genome Editor Delivery.Molecular biology · 2022Article
- Biocircuits in plants and eukaryotic algae.Frontiers in plant science · 2022Review
- Oral delivery of a functional algal-expressed TGF-β mimic halts colitis in a murine DSS model.Journal of biotechnology · 2021Article
21 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Microalgae have recently received attention as a potential low-cost host for the production of recombinant proteins and novel metabolites. However, a major obstacle to the development of algae as an industrial platform has been the poor expression of heterologous genes from the nuclear genome. Here we describe a nuclear expression strategy using the foot-and-mouth-disease-virus 2A self-cleavage peptide to transcriptionally fuse heterologous gene expression to antibiotic resistance in Chlamydomonas reinhardtii. We demonstrate that strains transformed with ble-2A-GFP are zeocin-resistant and accumulate high levels of GFP that is properly 'cleaved' at the FMDV 2A peptide resulting in monomeric, cytosolic GFP that is easily detectable by in-gel fluorescence analysis or fluorescent microscopy. Furthermore, we used our ble2A nuclear expression vector to engineer the heterologous expression of the industrial enzyme, xylanase. We demonstrate that linking xyn1 expression to ble2A expression on the same open reading frame led to a dramatic (~100-fold) increase in xylanase activity in cells lysates compared to the unlinked construct. Finally, by inserting an endogenous secretion signal between the ble2A and xyn1 coding regions, we were able to target monomeric xylanase for secretion. The novel microalgae nuclear expression strategy described here enables the selection of transgenic lines that are efficiently expressing the heterologous gene-of-interest and should prove valuable for basic research as well as algal biotechnology.
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