KONSTRUKSI GEN PENGKODE FARNESYL PYROPHOSPHATE SYNTHASE (FPS), SALAH SATU ENZIM PADA BIOSINTESIS OBAT ANTIMALARIA ARTEMISININ,KE DALAM VEKTOR BINER pCAMBIA 1303

• Main Author: Andam Dewi, Mutiara
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45493
• Institution: Institut Teknologi Bandung

Artemisinin is a secondary metabolite derived from Artemisia annua L. that is effective as anti-malarial drug. However, the level of artemisinin in A. annua is very low and artemisinin-based malaria treatment being relatively expensive. Therefore, an effective strategy to increase the levels of artemisinin is needed. Nowadays, researches thus have been developing leads to genetic engineering involving biosynthetic pathway of artemisinin. In artemisinin biosynthesis, farnesyl pyrophosphate synthase (FPS) is one of the enzymes which have an important role. This enzyme will be transform to A. annua in order to increase the level of artemisinin. To transforms FPS gene in to A. annua, this gene should be inserted to binary vector pCAMBIA 1303. This study aimedto insert FPS gene into binary vector pCAMBIA 1303. FPS gene used in this study is a synthetic gene that has been in the cloning vector pUC57. pUC57-FPS plasmid and empty pCAMBIA plasmid which has been reproduced were cut using restriction enzymes SpeI and NcoI. Construction was conducted by ligating two DNA fragments that have been truncated and subsequently transformed in Escherichia coli strain DH5?. The results showed that FPS gene was successfully inserted into a vector pCAMBIA 1303, so it produced a recombinant vector pCAMBIA 1303-FPS. Recombinant vector that has been isolated and analyzed by electrophoreses using 1% agarose gel and was confirmed by migration analysis, PCR using specific primers FPS, restriction analysis and determining the nucleotide sequence (sequencing). Confirmation results showed that recombinant plasmid was migrated slowly, PCR result showed there is a fragment DNA of FPS on the size of ±1032 bp and this fragment can be cut by using restriction enzyme XbaI. In the determination of FPS gene nucleotide sequence (sequencing), the result is not well-confirmed yet.