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. 2011 Jan 1;2(1):50–54. doi: 10.4161/bbug.2.1.13423

Figure 1.

Figure 1

Transformation of the Chlamydomonas reinhardtii chloroplast genome with genes encoding for human therapeutic proteins. Schematic diagram of the transformation vectors used and the corresponding integration sites. pD1-KanR: Replacement of the endogenous psbA gene with the gene of interest under the control of the psbA promoter and UTR elements. The kanamycin resistance gene aphA6 (KanR) under the control of the atpA promoter and 5′ UTR is genetically linked to the gene of interest. Grey regions flanking the gene of interest and resistance gene corresponds to regions of the chloroplast genome used for homologous recombination between the insertion plasmid and the C. reinhardtii chloroplast genome. pD1-KanR-SAA: Identical to pD1-KanR except that the gene of interest fused to the C-terminus of M-SAA. pAtpA: Vector used for the insertion of the genes of interest under the control of the atpA promoter and 5′ UTR and the rbcL 3′ UTR into the BamHI silent site between the psbA gene the 5S rRNA.19 All recombinant proteins were C-terminally fused to the 1 x FLAG-tag sequence (DYKDDDDKS) for western blotting and purification.