Fig. 1

Physical map of fine structure for the chloroplast specific DARPin G3 expression cassette (NT-ch DARPin G3 cassette), plastid transformation vectors of the pPRV111A harboring DARPin G3 expression cassettes (PRV-DARPin G3), targeting region in the wild-type tobacco plastid genome (Nt-chDNA) and transformed plastid genome regions (Nt-DARPin G3). Nt-Prrn: ribosomal RNA operon promoter from tobacco; T7g10 5′ UTR: 5′ untranslated region of bacteriophage T7 gene 10; DARPin G3: coding sequence of DARPin G3, TrrnB: rrnB 3′ untranslated region from E. coli; PpsbA: promoter and 5′ UTR of psbA gene; aadA: aminoglycoside 3′- adenylytransferase gene; TpsbA: terminator of psbA gene. The transgenes are targeted to the intergenic region between the rrn16 and rps7/12 plastid genes. Primers P1/P2 land on the DARPin G3 coding sequence, generating a 215 bp fragment. Primers P3/P4 land on the rrn16 and the rps7/12 flanking sequence, generating ~ 4 kb and ~ 2 kb fragments in transplastomic and wild-type plants, respectively. The expected sizes of the DNA fragments in Southern blot analyses with the restriction enzyme BglII are indicated. The location of the Southern blotting probe is shown as a black bar