KONSTRUKSI CYP71AV1, SALAH SATU GEN PADA BIOSINTESIS OBAT ANTIMALARIA ARTEMISININ, KEDALAM VEKTOR BINER pCAMBIA 1303

• Main Author: Julian, Wiji
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45479
• Institution: Institut Teknologi Bandung

Cytochrome P450 mono-oxygenase (cyp71av1) is an enzyme that has important function in biosynthesis of artemisinin in Artemisia annua L. Artemisinin is an effective drug for malaria treatment. However, the artemisinin content in A. annua L. is very low and causes the artemisinin-based therapy becomes relatively expensive. Nowadays, the main focus research in aiming to increase the production of artemisinin is using genetic transformation techniques. The objective of this study was to insert the cyp71av1 gene into the binary vector pCAMBIA 1303. cyp71av1 gene that previously has been cloned in pJET1.2/Blunt then amplified with Polymere Chain Reaction (PCR) using specific cyp71av1 primer which has BglII and SpeI restriction site. Construction is done by ligating pCAMBIA 1303 and cyp71av1 that had been digested by restriction enzymes BglII and SpeI. The results showed that the cyp71av1 gene has been successfully inserted into pCAMBIA 1303, resulting in pCAMBIA-cyp71av1 recombinant vector. The isolated recombinant vector was confirmed by PCR using the specific cyp71av1 primer, and gusA (a reporter gene from pCAMBIA 1303) primer, and DNA sequencing. The confirmation using PCR result showed that there was a cyp71av1 DNA fragment in ±1500 bp band. The PCR result using the gusA primer also showed the gusA band. However, the cyp71av1 sequencing results is not wellconfirmed yet, so it needs to be optlimalized again.