TRANSFORMATION OF 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase-1 (hmgr1) GENE MEDIATED BY Agrobacterium tumefaciens OF Andrographis paniculata TO IMPROVE ANDROGRAPHOLIDE CONTENT
<p align="justify">HMGR (3-Hydroxy-3-methylglutaryl Coenzyme A Reductase) is one of the enzymes involved in biosynthesis of andrographolide, diterpene lactone. HMGR catalyzed the reaction on the andrographolid synthesis pathway from HMG-CoA (3-Hydroxy-3-methylglutaryl coenzyme A) to...
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| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/28393 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | <p align="justify">HMGR (3-Hydroxy-3-methylglutaryl Coenzyme A Reductase) is one of the enzymes involved in biosynthesis of andrographolide, diterpene lactone. HMGR catalyzed the reaction on the andrographolid synthesis pathway from HMG-CoA (3-Hydroxy-3-methylglutaryl coenzyme A) to mevalonate. Our previous study indicated that production of andrographollid demonstrated limited amount. On the other hand, several reports indicated that genetically engineered plant cell culture showed a potency for enhancing secondary metabolites production. One such modification can be done by introducing a gene of interest into plant cells to increase the activity of specific enzymes which regulated the synthesis of desired secondary metabolites. The purposed of the study was to transformed cotyledone of Andrographis paniculata with hmgr1 gene mediated by Agrobacterium tumefaciens GV3101 strains that had been constructed with recombinant plasmid pBI121, regenerated transformed A. paniculata tissue, and evaluated the andrographolid content by the High Performance Liquid Chromatography (HPLC) method at the transformed plant age of three months. The marker gene in A. paniculata used for selection was kanamycin at concentration of 20 ppm. Analysis of PCR (Poly Chain Reaction) indicated the presence of 2.262 bp fragment that confirmed pBI121-hmgr1. This confirmed that the hmgr1gene has been integrated into the DNA genome of A. paniculata tissue. The result of andrografolid content in the transformed plant analysis using HPLC method was 3.63% and the andrografolid content in the control plant was 2.94%. This result indicated that the insertion of hmgr1 gene in A. paniculata could increase the andrographolide content by 23.5%. <p align="justify"> |
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