Expression of OfCCD1 in b-carotene accumulating
cells. The cDNA was expressed in E. coli XL1Blue
harboring the pACCAR plasmid that carries all genes
necessary to produce β-carotene (Missawa et al., 1995).
A single colony of transformed cells was inoculated in
100 mL YT media containing chloramphenicol and carbenicillin
and grown at 37°C over night. One mL overnight
culture was inoculated in 100 mL of fresh YT media
containing the suitable antibiotics and grown at 27°C
until an OD550 of 0.5. To this culture ITPG (0.5 mM),
ascorbate (6 mM), FeSO4 (5 μM), catalase (200 U/mL)
were added to induce the production of the target enzyme
and the in vivo reaction.