Development of a Safer Platform for the Production of Recombinant Product than GM Crops

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Many genetically modified (GM) crops were used for production of plant-derived edible vaccines and other therapeutic recombinant products. However, GM crops resulted in the ecotoxicological risk of gene transfer because of pollen flow during the past 20 years. The most-commonly used eukaryotic model alga, Chlamydomonas reinhardtii has recently been shown the potential of decreasing this risk. Compared to GM crops, there is no risk of gene transfer because the alga culture can be deserved completely scrutiny under laboratory condition and it do not produce pollen. Recently, we had transformed the chloroplast of Chlamydomonas reinhardtii with two genes, CTB and CV1, which encode cholera toxin B subunit and chimeric antigen CV1 fused CTB with VP1 protein from foot and mouth disease virus (FDMV). The transgenic alga subculture were carried out under different selective conditions. The recombinant antigen in transgenic Chlamydomonas chloroplast was detected by western blotting in a period of subculture time. However, the PCR detection data demonstrated that transgene integrated with chloroplast genome would be lost in a special time when was connected with subculture condition. Although loss of transgenic fragment was an inevitable fate for the green alga, our research data showed the possibility that the presence of transgenic fragment was strictly regulated. Thus, the alga can be used for a safer platform for the production of recombinant product than GM crops.

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424-429

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January 2013

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© 2013 Trans Tech Publications Ltd. All Rights Reserved

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[1] T. Cardi, P. Lenzi, P. Maliga (2010) Chloroplasts as expression platforms for plant-produced vaccines. Expert Rev Vaccines 9: 893–911.

DOI: 10.1586/erv.10.78

Google Scholar

[2] S. J. Stratified (2007) Approaches to achieve high-level heterologous protein production in plants. Plant Biotechnol J 5: 2-15.

Google Scholar

[3] B. A. Rasala, S. P. Mayfield (2011) The microalga Chlamydomonas reinhardtii as a platform for the production of human protein therapeutics. Bioeng Bugs 2: 50-54.

DOI: 10.4161/bbug.2.1.13423

Google Scholar

[4] S. P. Mayfield, et al (2007) Chlamydomonas reinhardtii chloroplasts as protein factories. Curr Opin Biotechnol 18: 126-133.

Google Scholar

[5] B. A. Rasala, et al (2010) Production of therapeutic proteins in algae, analysis of expression of seven human proteins in the chloroplast of Chlamydomonas reinhardtii. Plant Biotechnol J 8: 719–733.

DOI: 10.1111/j.1467-7652.2010.00503.x

Google Scholar

[6] R. Surzycki, et al (2009) Factors effecting expression of vaccines in microalgae. Biologicals 37: 133–138.

DOI: 10.1016/j.biologicals.2009.02.005

Google Scholar

[7] A. Coragliotti, et al (2011) Molecular factors affecting the accumulation of recombinant proteins in the Chlamydomonas reinhardtii chloroplast. Mol Biotechnol 48: 60-75.

DOI: 10.1007/s12033-010-9348-4

Google Scholar

[8] C. M. Hutchinsa (2010) Transcriptomic signatures in Chlamydomonas reinhardtii as Cd biomarkers in metal mixtures. Aquat Toxicol 100: 120–127.

DOI: 10.1016/j.aquatox.2010.07.017

Google Scholar

[9] D. Tolleter, et al (2011) Control of hydrogen photoproduction by the proton gradient generated by cyclic electron flow in Chlamydomonas reinhardtii. Plant Cell 23 : 2619-2630.

DOI: 10.1105/tpc.111.086876

Google Scholar

[10] S. Mayfield, S. Franklin, R. Lerner (2003) Expression and assembly of a fully active antibody in algae. Proc Nat Acad Sci 100: 438-442.

DOI: 10.1073/pnas.0237108100

Google Scholar

[11] B. A. Rasala, et al (2012) Robust Expression and Secretion of Xylanase1 in Chlamydomonas reinhardtii by Fusion to a Selection Gene and Processing with the FMDV 2A Peptide. PLoS One 7 : e43349.

DOI: 10.1371/journal.pone.0043349

Google Scholar

[12] S. Rosales-Mendoza, L. M. Paz-Maldonado, R. E. Soria-Guerra. (2012) Chlamydomonas reinhardtii as a viable platform for the production of recombinant proteins: current status and perspectives. Plant Cell Rep 31: 479-94.

DOI: 10.1007/s00299-011-1186-8

Google Scholar

[13] X. Wang, et al (2008) A novel expression platform for the production of diabetes-associated autoantigen human glutamic acid decarboxylase (hGAD65). BMC Biotechnol 8: 87.

DOI: 10.1186/1472-6750-8-87

Google Scholar

[14] E. S. Alke, et al (2009) Strategies to facilitate transgene expression in Chlamydomonas reinhardtii. Planta 229: 873-83.

DOI: 10.1007/s00425-008-0879-x

Google Scholar

[15] J. A. Gregory, et al (2012) Algae-produced Pfs25 elicits antibodies that inhibit malaria transmission. PLoS One 7: e37179.

DOI: 10.1371/journal.pone.0037179

Google Scholar

[16] E. Specht, M. S. Shigeki, S. P. Mayfield (2010) Micro-algae come of age as a platform for recombinant protein production. Biotechnol Lett 32: 1373–1383.

DOI: 10.1007/s10529-010-0326-5

Google Scholar

[17] A. L. Manuell, et al (2007) Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast. Plant Biotechnol J 5: 402-412.

Google Scholar

[18] L. Michelet, et al (2011) Enhanced chloroplast transgene expression in a nuclear mutant of Chlamydomonas. Plant Biotechnol J 9: 565-574.

DOI: 10.1111/j.1467-7652.2010.00564.x

Google Scholar

[19] J. E. Maul, et al (2002) The Chlamydomonas reinhardtii plastid chromosome: islands of genes in a sea of repeats. Plant Cell 14: 2659-2679.

DOI: 10.1105/tpc.006155

Google Scholar

[20] A.R. Grossman, et al (2003) Chlamydomonas reinhardtii at the crossroads of genomics. Eukaryot Cell 2: 1137-1150.

Google Scholar

[21] D. Dauvillee, et al (2010) Engineering the chloroplast targeted malarial vaccine antigens in Chlamydomonas starch granules. PLoS ONE 5: e15424.

DOI: 10.1371/journal.pone.0015424

Google Scholar