Evidence for T-DNA mediated gene targeting to tobacco chloroplasts.

The integration of foreign DNA into plant cells by Agrobacterium mediated transformation is random and normally directed at the nucleus. Here we present evidence that such transformation can be used to introduce foreign genes into higher plant chloroplasts by site-specific homologous recombination. A binary vector was made in vitro that included chloroplast ribosomal DNA (rDNA) within T-DNA borders, and transgenic tobacco plants obtained using this construct were analyzed for the targeted recombination by hybridization, polymerase chain reaction (PCR) and DNA sequencing.