Southern blot analysis. A flanking sequence probe (810 bp) was obtained from the trnA/trnI genes (indicated by red lines in 4A). Regulatory and coding sequences are the same as in figure 3A. This map also shows the chloroplast DNA fragment that hybridizes with this probe. Untransformed genome hybridized with a 4.47 kbp fragment whereas transformed chloroplast genomes hybridized with two fragments of 5.2 kbp and 930 bp. B) Lane 1: Untransformed control; Lanes 2–3: IGF-1s transgenic lines (T0 generation); Lanes 4–5: IGF-1s transgenic lines (T1 generation); Lanes 6–7: IGF-1n plants (T0 generation); Lane 8: positive control. C) The IGF-1 coding sequence was used as a probe to confirm integration of the IGF-1 gene into the chloroplast genome. The transgenic lines that contain the IGF-1 show a 930 bp fragment. Lane: 1- untransformed, Lanes 2–3: IGF-1s transgenic lines (T0), 4–5: IGF-1s transgenic lines (T1); 6–7: IGF-1n transgenic lines; Lane 8: blank; Lane 9: the IGF-1 probe as a positive control.