PENYISIPAN GEN ARTEMISINIC ALDEHYDE ?11(13) DOUBLE BOND REDUCTASE (DBR2) KE DALAM PLASMID pCAMBIA1303 YANG MENGANDUNG GEN SILENCING SUPRESSOR (p19)

• Main Author: Haniffadli, Ariranur
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44353
• Institution: Institut Teknologi Bandung

Artemisinic a?¸±í?¸± ???????double bond reductase (DBR2) was one of enzymes that responsible to catalyze the dihydroartemisinic aldehide as precursor for artemisnin biosynthesis. DBR2 encoded gene was transformed to Artemisia annua L. plant for increasing of antimalaria artemisinin production. Agroinfiltration was a method that can be used for transformation of extraneous gene into plant. However, this transformation can activate defense system of plant with non-activated extraneous gene that entered the plant called post transcriptional gene silencing (PTGS) mechanism. This mechanism caused transformant gene can not be translated by plant so that the expression of the enzyme was blocked. Some virus can produce protein that can disturbe the defense system of plant (silencing supressor) such as p19 protein. P19 protein is derived from tomato bushy stunt virus (TBSV) that coded by p19 gene. Therefore, the objective of this research was insertion of DBR2 gene into the p19 containing pCAMBIA1303 plasmid, that can be transformated into A. annua L.. The method used for insertion was cut and paste of synthetic DBR2 gene to pCAMBIA1303 plasmid which contain p19 gene. DBR2 gene synthetic was restricted by BglII enzyme and SpeI enzyme then ligated into the p19 containing pCAMBIA1303 plasmid. Recombinant plasmid has been analysed with migration, PCR, restriction and sequencing. Based on allignment of sequence data, it could be concluded that the DBR2 gene was successfully inserted into p19 containing pCAMBIA1303 plasmid with query coverage 55% and identical 100%.