Fig. 1.

Transformation of Synechococcus PCC7002 bicA into the inverted repeat (IR) regions of the tobacco plastome. (A) Tobacco plastome sequence in transforming plasmid pRV112a ((Zoubenko et al., 1994); cloned IR_B sequence regions are indicated. rps16 3′-UTR (t) and rrn promoter and 5′-UTR (p) directed integration of the selectable marker genes aadA and bicA by homologous recombination (Whitney and Sharwood, 2008). The tobacco psbA promoter/5′-UTR (P) and 114-bp of its 3′-UTR (T) sequence were used to regulate expression of bicA modified to code 10 N-terminal residues of the Rubisco L-subunit located 5′ to a unique NheI cloning site. Correct transformation into the tobacco plastome introduced a unique HindIII site (H). The annealing positions of nucleotide probes are shown. (B) Total leaf DNA from independently transformed T₁ tob^BicA lines examined by Southern blot analysis. In wild-type plants (wt), the ³²P-labelled 900-bp TGF probe recognizes a single 7.7-kb fragment and a 3.8-kb fragment in the transformants.