Expression Of Candida SLS Candidate Gene In Tobacco

Camptotheca acuminata Decne is endemic species of China. It contains indolylalkylamine alkaloid which is used as anti-cancerous chemicals. Its biosynthetic pathway involves many emzymes among which Secologanin synthase is coded by sls gene. Our laboratory has cloned a gene which has similar fonctions to sls gene, which is called sls-like gene. In this work, the open reading frame of sls-like gene is expressed in E.coli DH5a after it was inserted into plasmid pBI121. Then, the recombination of this gene in the plasmid pBI121 (called pBI190-1575) has been conformed by PCR, electrophoresis and DNA sequence. The sls-like gene was transfered to tobacco via Agrobacterium tumefaciens vector. Transgenic plants were obtained in selective medium containing kanamycin. Putatively transformed tobacco leaves were verified by bud regeneration and PCR. Protein profiles were compared between the two plasmids (pBI190-1575, pBI121), and between transgenic and non-transgenic tobacco. Our results showed that sls-like gene has been integrated into prokaryotic(E.coli) and eukaryotic cells (tobacco).