Methods: Engineered T-DNA
- Three Ti-plasmids were
constructed with the following T-DNA sequences
- psy,
crtI, and lcy - corresponding enzymes of the b-carotene
biosynthetic pathway
- Gt1
p & 35S p - endosperm-specific glutelin and constitutive
cauliflower mosaic virus 35S promoters
- aphIV
& nptII - selectable markers (hygromycin & kanamycin-resistence
respectively)
- nos!
& 35S! - polyadenylation signals (necessary for functional
eucaryotic transcripts)
- tp
- transit peptide sequence (translocation of gene products to
the plastids of the endosperm)
- I-sce-I, Kpn I & Spe I - restriction sites
- RB & LB - right & left
border sequences (necessary for proper T-DNA insertion)
- Two transformation experiments
were attempted
- pB19hpc single-transformants -
800 rice embryos were infected with A. tumefaciens containing
the pB19hpc plasmid.
- pZPsC/pZLcyH co-transformants -
500 rice embryos were infected with a mixture of A. tumefaciens
containing the pZPsC plasmid and A. tumefaciens containing
the pZLcyH plasmid.
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