Regulation of camptothecin in plant tissue cultures: Construction of the recombinant fusion strictosidine synthase-GFP in pJIT60

Strictosidine is an important intermediate in the formation of over 3000 alkaloids including camptothecin (Hallard et al., 1998). In this project, the strictosidine synthase gene was fused to the gene for the green fluorescent protein marker in the plasmid pEGFP-N1, obtained from Clontech, Inc. This fusion fragment containing both the strictosidine synthase gene and the green fluorescent protein gene was excised from the pEGFP-N1 plasmid and introduced into the plant expression vector pJIT60. Studies were carried out to determine green fluorescence and strictosidine synthase activity in mammalian cells as well as in the Camptotheca acuminata plant cells. The primary goal of this thesis research was to construct a recombinant vector, not yet commercially available, which could be used to study the regulation of camptothecin in plant tissue cultures, by way of understanding the formation of strictosidine.