Transgenic Tobacco Expressing The Crocus CSzCD Gene Showed The Production Of Crocetin And Study On The Anticancer Effects Of Chemosynthesis Crocetin In Vitro

Saffron (Crocus sativus L.), a perennial herbage of Iridaceae, is a preciousherbal medicine and widely used as a colorant and flavoring agent. The stigma is themedica part, including crocetin, crocin, picrocin and safranal. To explore theresources of active ingredients from the saffron and further investigate systemicly theeffects of crocetin on the growth inhibition in carcinoma cell lines. In this research,the CSzCD gene was cloned from the saffron and transformed into the tobacco plant,the purpose was to build the bioreactor which produced the crocetin by the transgenictobacco plants. Meanwhile we conducted the systemic anti-tumor research tochemosynthesis crocetin in vitro.To investigate the feasibility to produce crocetin in tobacco plants. In this study,the coding region of zeaxanthin cleavage dioxygenase (CSzCD) gene from crocussativus L. was inserted into the downstream of the caμliflower mosaic virus (CaMV)35S promoter of a binary vector pBI121, and integrated into the genome of Nicotianatabacum L. Twenty-one transgenic lines were identified by genomic southernblotting analysis. Western blotting and HPLC analysis of the leaf extracts oftransgenic tobacco showed that crocetin was produced in CSzCD gene-transformedplants, while no crocetin was found in the untransformed tobacco. To observe the effects of the chemosynthesis crocetin on the growth inhibitionin five carcinoma cell lines. Methods. the growth inhibition was analyzed by MTT,DNA Ladder and cell growth curve measurement technique in Raji,K562,A549,CoCa2 and BIU-87 cells lines.