MOLECULAR CLONING AND DESIGNING TRANSGENIC CONSTRUCT CARYING FLAVONOID 3’ 5’ HYDROXYLASE GENE ISOLATED FROM Aconitum carmichaelii Debx. PLANTS

Abstract

Aconitum carmichaelii is a medicinal plant which contains many valuable pharmaceutical substances, including flavonoids. Flavonoids are important compounds that have antioxidant, anti-cell proliferative and especially anti-cancer activities, however flavonoid content in A. carmichaelii plant is very low. Flavonoid 3 '5' hydroxylase (F3'5'H) is an important enzyme that catalyzes the final reaction in flavonoid biosynthesis in A. carmichaelii plants. Therefore, this studying approach of overexpression of gene encoding F3’5’H in order to improve flavonoid content is an effective strategy in research of medicinal plants. In this study, the authors present the characteristics of Aconitum carmichaelii flavonoid 3 '5' hydroxylase (AcF3'5'H) gene isolated from mRNA of A. carmichaelii plant, and the design of construct carrying AcF3'5'H transgene in plant expression vector to enhance AcF3'5'H gene expression in transgenic A. carmichaelii Debx plants. The coding segment of AcF3'5'H gene has 1521 bp, which encodes 506 amino acids. The construct carrying AcF3'5'H transgene in pCB301 gene transfer vector contains 35S promoter, c-myc and KDEL sequences. The pCB301_AcF3’5’H gene transfer vector was transformed into Agrobacterium tumefaciens to produce recombinant bacteria.