Figure 1

DNA gene constructs, site of integration and identification of transformants in Chlamydomonas reinhardtii chloroplast. a Transformation vector containing the genes of interest: Gluc (Luciferase from Gaussia princeps), or GFP (Green Fluorescent Protein from Aequorea vcitoria), or Immunotoxin (αCD22CH23PE40 (Tran et al. 2013b)). Regions of the chloroplast genome were positioned at both ends of the transformation vector to allow homologous integration of the whole transformation cassette into the chloroplast genome (w1.1). b, c Colony PCR amplification to confirm the presence of recombinant genes of interest, Gluc—Luciferase from Gaussia princeps (b) or GFP—Green Fluorescent Protein (c). b, c Lanes 1, 2 and 3 show three different gene positive colonies (dotted arrow). d, e Colony PCR amplification to check homoplasmicity by using primers from the genome to verify the absence of the second band (dashed arrow), which indicates the replacement of SAA by our recombinant genes. The black arrow shows the amplified control PCR product. d, e Lanes 1 and 3 show homoplasmic colonies, and lane 2 a control. All the arrows (dotted, dashed and black) are also marked (a) to indicate the gene amplified regions.