PENYISIPAN GEN P19 KE DALAM VEKTOR REKOMBINAN pCAMBIA-FPS DALAM UPAYA MENINGKATKAN EFISIENSI TRANFORMASI GEN FPS KE TANAMAN ARTEMISIA ANNUA L.

• Main Author: Nurur Rohmah, Mulyani
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44365
• Institution: Institut Teknologi Bandung

Nowadays, treatment of malaria caused by Plasmodium falciparum according to WHO recommendation is by artemisinin based combination therapy. Artemisinin is one of secondary metabolites in Artemisia annua L. for treating malaria. Naturally, its concentration is low and this condition influencing the cost for treating malaria by artemisinin combination therapy become expensive. Attempt to synthesis its compound is hard to do. So that, the strategy that used to increase artemisinin concentration in plant is by genetic engineering. Genes encoded key enzyme in artemisinin biosynthetic patways have been found and investigated. FPS or farnesyl pyrophosphate synthase is one of key enzyme which catalize reaction to produce farnesyl pyrophosphate. Farnesyl pyrophosphate is a precursor in production of isoprenoid compound, one of them is artemisinin. Overexpression of fps gene in A. annua L. have been done by scientists. However, the concentration of artemisinin produced is only 1 % of dry weight. The presence of post transcriptional gene silencing as a protection mechanism in plant is assumed as the reason of the transformation of the gene into the plant is less effective, so that the artemisinin produced is low. This study aimed to insert antisilencing p19 gene into vector containing fps gene, pCAMBIA- FPS, in order to antisilince the plant protection mechanism when the plasmid transformed in to the plant. Strategy that used to construct pCAMBIA-FPS-p19 plasmid was by restricting pCAMBIA-p19 plasmid by Xhol restriction enzyme as an insert gene, and restricting pCAMBIA-FPS by Xhol restriction enzyme, as a vector. Insert gene and vector fragment is ligated using T4 ligase enzyme then the product from ligation is transformed to E.coli Dh5????. Based on the analysis of migration, PCR, restriction and sequencing, it could be concluded that the p19 gene has been succesfully inserted into pCAMBIA-fps plasmid.