TRANSFORMASI GEN AMORFA-4,11-DIEN SINTASE (ADS) KE DALAM KULTUR IN VITRO ARTEMISIA ANNUA L. MENGGUNAKAN AGROBACTERIUM TUMEFACIENS AGL-1 UNTUK MENINGKATKAN KADAR ARTEMISININ

• Main Author: Chamidah, Nur
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/64758
• Institution: Institut Teknologi Bandung

Artemisinin, isolated from Artemisia annua L. is a sesquiterpene lactone endoperoxide and used as an antimalarial drug. The artemisinin content in this plant is quite low, so enhancement of its content is needed through genetic transformation and the best transformation condition was determined at 0D600 0,5; 0,8; and 1,0. Transformation of A. annua in vitro culture was mediated by Agrobacterium twnefaciens AGL-1 with pCAMBIA 303-ads vector using vacuum infiltration method to obtain the highest frequency of transformation. The results were analyzed based on transient of p¬glucuronidase (GUS) expression and the artemisinin content were measured using high performance liquid chromatography (HPLC). The best transformation condition was obtained at 0D600 1.0 with the frequency of transformation was 80.61 0.02 % while the frequency of transformation at 0D600 0.5 and 0.8 was 47.86 ± 0.26 % and 69.59 ± 0.14 %. The artemisinin content increased more than the untransformed culture. Transformation performed at OD600 0.5, 0.8, and 1.0 respectively produced 3.78 + 0.22; 6.14 0.24, and 7.49 ± 0.34 ug/g extract of artemisinin while the artemisinin content in untransformed culture was 2.30 ± 0.31 ug/g extract of artemisinin. The transformed culture at 0D600 1.0 produced artemisinin 3.25 times higher than the untransfonned culture.