Isolation and Characterization of DBR2 Gene Promoter from Iranian Artemisia annua

Authors
R. Sarvestani
S. A. Peyghambary
A. Abbasi
Department of Agronomy and Plant Breeding, Agricultural College, University of Tehran, Karaj, Islamic Republic of Iran.
Abstract
Artemisia annua is still the only commercial source of Artemisinin. To date, a number of biochemical and molecular studies about Artemisinin’s biosynthetic pathway have been carried out. In metabolic engineering approach, isolation and characterization of promoters leads to an understanding of which cis-acting elements are responsible for the regulation of gene expression. DBR2 is a key enzyme in Artemisinin biosynthetic pathway. In order to allow chromosome walking beyond the 5'-flanking region of DBR2, two specific primers were used in combination with 6 arbitrary primers in TAIL-PCR method. A 696bp upstream of DBR2 start codon was isolated and cloned. The subsequent sequence analysis using bioinformatics softwares revealed that there were several cis-acting elements such as TATA-box, CAAT-box, and MeJA-responsive element, and several W-box and light-responsive elements inside the DBR2 promoter. These results can be helpful in understanding of artemisinin biosynthesis regulation and will facilitate metabolic engineering of the compound.

Keywords

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Journal of Agricultural Science and Technology
Volume 16, Issue 1
January and February 2014
Pages 191-202