Enhanced chloroplast transgene expression in a nuclear mutant of Chlamydomonas

Article Article scientifique Article Enhanced chloroplast transgene expression in a nuclear mutant of Chlamydomonas eng 2011 author Michelet Laure 663791 author Legendre Lefebvre Linnka 134661 author Burr Sarah E author Rochaix Jean-David 25289 author Goldschmidt-Clermont Michel P. 25594 565-574 10.1111/j.1467-7652.2010.00564.x 20809927 1467-7644 Chloroplast transformation in microalgae offers great promise for the production of proteins of pharmaceutical interest or for the development of novel biofuels. For many applications, high level expression of transgenes is desirable. We have transformed the chloroplast of Chlamydomonas reinhardtii with two genes, acrV and vapA, which encode antigens from the fish pathogen Aeromonas salmonicida. The promoters and 5' untranslated regions of four chloroplast genes were compared for their ability to drive expression of the bacterial genes. The highest levels of expression were obtained when they were placed under the control of the cis-acting elements from the psaA-exon1 gene. The expression of these chimeric genes was further increased when a nuclear mutation that affects a factor involved in psaA splicing was introduced in the genetic background of the chloroplast transformants. Accumulation of both the chimeric mRNAs and the recombinant proteins was dramatically increased, indicating that negative feedback loops limit the expression of chloroplast transgenes. Our results demonstrate the potential of manipulating anterograde signalling to alter negative regulatory feedback loops in the chloroplast and improve transgene expression. Aeromonas salmonicida/genetics/immunology Antigens, Bacterial/genetics Cell Nucleus/genetics Chlamydomonas reinhardtii/genetics/metabolism Chloroplasts/genetics/metabolism Gene Expression Regulation, Bacterial Gene Expression Regulation, Plant Genetic Engineering/methods Mutation Plants, Genetically Modified/genetics/metabolism Recombinant Proteins/biosynthesis/genetics Transformation, Genetic Transgenes Dewey 570 Dewey 580 43 Département de biologie moléculaire 161 45 Département des sciences végétales 163 false Plant biotechnology journal 9 5 22447bef-61b1-402f-bf03-ee7207a178b0 Article (Accepted version) Public Michelet et al 2010.pdf 1232702 application/pdf openAccess Faculté des sciences / Section de biologie / Département de biologie moléculaire Faculté des sciences / Section de biologie / Département des sciences végétales

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Enhanced chloroplast transgene expression in a nuclear mutant of Chlamydomonas	Plant biotechnology journal			2011	724	1,225

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Michelet, Laure