ArticleBMC biotechnology2008
A novel expression platform for the production of diabetes-associated autoantigen human glutamic acid decarboxylase (hGAD65).
Article in BMC biotechnology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Harnessing the potential of microalgae for the production of monoclonal antibodies and other recombinant proteins.Protoplasma · 2024Review
- Inhibition of NK cell cytotoxicity by tubular epithelial cell expression of Clr-b and Clr-f.Current research in immunology · 2024Article
- Chloroplast Engineering: Fundamental Insights and Its Application in Amelioration of Environmental Stress.Applied biochemistry and biotechnology · 2023Review
- Review
- Current Status and Perspective on the Use of Viral-Based Vectors in Eukaryotic Microalgae.Marine drugs · 2022Review
- Current advances and future prospects in production of recombinant insulin and other proteins to treat diabetes mellitus.Biotechnology letters · 2022Review
- Harnessing the Algal Chloroplast for Heterologous Protein Production.Microorganisms · 2022Review
- Plastid Transformation: How Does it Work? Can it Be Applied to Crops? What Can it Offer?International journal of molecular sciences · 2020Review
- Translocation from the chloroplast stroma into the thylakoid lumen allows expression of recombinant epidermal growth factor in transplastomic tobacco plants.Transgenic research · 2020Article
- Review
- Intercistronic expression elements (IEE) from the chloroplast of Chlamydomonas reinhardtii can be used for the expression of foreign genes in synthetic operons.Plant molecular biology · 2018Article
- Recent achievements obtained by chloroplast transformation.Plant methods · 2017Review
- The Potential for Microalgae as Bioreactors to Produce Pharmaceuticals.International journal of molecular sciences · 2016Review
- Development of phytase-expressing chlamydomonas reinhardtii for monogastric animal nutrition.BMC biotechnology · 2016Article
- Transgene Expression in Microalgae-From Tools to Applications.Frontiers in plant science · 2016Review
- Photosynthetic biomanufacturing in green algae; production of recombinant proteins for industrial, nutritional, and medical uses.Photosynthesis research · 2015Review
- Recent advances on host plants and expression cassettes' structure and function in plant molecular pharming.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2014Review
- Algae-based oral recombinant vaccines.Frontiers in microbiology · 2014Review
- Production of therapeutic proteins in the chloroplast of Chlamydomonas reinhardtii.AMB Express · 2014Article
- Comparative evaluation of recombinant protein production in different biofactories: the green perspective.BioMed research international · 2014Review
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7 authors.
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Abstract
backgroundHuman glutamic acid decarboxylase 65 (hGAD65) is a key autoantigen in type 1 diabetes, having much potential as an important marker for the prediction and diagnosis of type 1 diabetes, and for the development of novel antigen-specific therapies for the treatment of type 1 diabetes. However, recombinant production of hGAD65 using conventional bacterial or mammalian cell culture-based expression systems or nuclear transformed plants is limited by low yield and low efficiency. Chloroplast transformation of the unicellular eukaryotic alga Chlamydomonas reinhardtii may offer a potential solution.
resultsA DNA cassette encoding full-length hGAD65, under the control of the C. reinhardtii chloroplast rbcL promoter and 5'- and 3'-UTRs, was constructed and introduced into the chloroplast genome of C. reinhardtii by particle bombardment. Integration of hGAD65 DNA into the algal chloroplast genome was confirmed by PCR. Transcriptional expression of hGAD65 was demonstrated by RT-PCR. Immunoblotting verified the expression and accumulation of the recombinant protein. The antigenicity of algal-derived hGAD65 was demonstrated with its immunoreactivity to diabetic sera by ELISA and by its ability to induce proliferation of spleen cells from NOD mice. Recombinant hGAD65 accumulated in transgenic algae, accounts for approximately 0.25-0.3% of its total soluble protein.
conclusionOur results demonstrate the potential value of C. reinhardtii chloroplasts as a novel platform for rapid mass production of immunologically active hGAD65. This demonstration opens the future possibility for using algal chloroplasts as novel bioreactors for the production of many other biologically active mammalian therapeutic proteins.
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