KONSTRUKSI ARTEMISINIC ALDEHYDE ?11(13) DOUBLE BOND REDUCTASE (DBR2), SALAH SATU GEN YANG BERPERAN PADA BIOSINTESIS OBAT ANTIMALARIA ARTEMISININ, KE DALAM VEKTOR BINER pCAMBIA 1303

• Main Author: Khairunisa, Masmaranti
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45482
• Institution: Institut Teknologi Bandung

Malaria is a serious parasitic infection disease in medical field. Aminoquinolin has been used as the main choice to treat malaria, but resistence from the microbe to this drug occurs gradually so that the more effective therapy for malaria is needed. In 2005, WHO recommends treatment of malaria with artemisinin-based combination called ACTs (artemisinin-based combined therapies) as the first choice in the treatment of malaria cases. However, the levels of artemisinin in the plant Artemisia annua L. very low, so that efforts to increase the production of artemisinin via genetic transformation for the important enzymes into a major research have been developed. Artemisinic aldehyde ?11 (13) reductase (DBR2) is one of the enzymes which have an important role in the biosynthesis of artemisinin. This study aimed to insert artemisinic aldehyde ?11 (13) reductase (DBR2) gene on the binary vector pCAMBIA 1303. First step was design synthetic gene for DBR2 by adding the restriction side BglII and SpeI at the gene sequence of DBR2. DBR2 gene that has been inserted into vector pUC57 was then transferred into the binary vector pCAMBIA 1303 by ligation based method. Before ligation, binary vector pCAMBIA 1303 and gene DBR2 each cut using restriction enzymes BglII and SpeI. The ligation results were transformed into E.coli DH5?. The recombinant plasmid obtained from the transformation was then confirmed by migration and restriction analysis, PCR product with gene-specific primer DBR2, and determination of DNA sequence. The results showed that the gene DBR2 has been successfully inserted into binary vector pCAMBIA 1303 and resulted the recombinant plasmid pCAMBIA 1303-DBR2. It was confirmed by the migration site of the recombinant plasmid pCAMBIA 1303-DBR2 shorten than plasmid pCAMBIA 1303 and pUC57-DBR2. The cutted pCAMBIA 1303-DBR2 and cutted empty pCAMBIA 1303 showed different sizes result. The PCR product showed the presence of the gene band at ± 1100 bp which is the size of the gene DBR2. However, the DBR2 sequencing results is not well-confirmed yet, so it needs to be optimalized again.