Construction of chloroplast transformation vector and its functional evaluation in L.

Chloroplast transformation vectors require an expression cassette flanked by homologous plastid sequences to drive plastome recombination. The 16-23 plastome region was selected and using this region, a new species-specific plastid transformation vector CuIA was developed with pKSII as a backbone by inserting the 16- and -23 sequences from L. An independent expression cassette with gene encoding aminoglycoside 3'-adenylyltransferase with controlling elements is added into the - intergenic region that confers resistance to spectinomycin. An efficient plastid transformation in bitter melon ( L.) was achieved by bombardment of petiole segments. The frequency of transplastomic plants yielded using standardized biolistic parameters with CuIA vector was two per 15 bombarded plates, each containing 20 petiole explants. Integration of gene was verified by PCR analysis in transplastomes. Transplastomic technology developed may be a novel approach for high level expression of pharmaceutical traits.