Figure 5: (A ) Illustration of the plastome locus downstream of psbA prior to insertion of the mVenCP andcodA-aadA cassettes (strain Luc2), as the intermediate recombinant locus (R1), and following loop-out of the codA-aadAmarker (R2). The locations of the primers (P1 – P4) used in a four-primer PCR analysis of transformant lines are indicated together with the expected product sizes. (B ) PCR conformation of homoplasmy at the R2 stage for four independent Luc2:Ven1 transformant lines. The Luc2 control showed the expected WT bands at 2.5 kb, whereas all four transformants showed the expected R2 PCR product at 1.5 kb. (C ) Spot tests of mid-log cultures, and 1:10 and 1:100 dilutions, of WT, Luc2 and Luc2:Ven1 on TAP plates containing: no antibiotic, Spc or 5-FC. (D ) Fluorescence analysis of Luc2:Ven1 compare to WT. Fluorescence measurements were taken on OD750 normalized mid-log cultures as three replicates (3n). (E ) Microplate-based relative luminescence analysis of WT, Luc2 and Luc2:Ven1. Luminescence measurements were taken on OD750 normalized mid-log cultures as three replicates (3n).