PENGARUH PENAMBAHAN GEN P19 PADA PLASMID YANG MENGANDUNG GEN ADS TERHADAP EFISIENSI TRANSFORMASI DENGAN TEKNIK EKSPRESI TRANSIEN BERBASIS AGROBACTERIUM PADA KULTUR DAUN ARTEMISIA ANNUA L. MELALUI ANALISIS HISTOKIMIA
Malaria is still remain a serious problem in the world. In Southeast Asia, malaria reached 28 million cases by 2010. Artemisinin is a secondary matebolite produced by Artemisia annua L. which is used as a first choice drug in form of ACTs (artemisinin-based combined therapies) to treat malaria as re...
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| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/64754 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Malaria is still remain a serious problem in the world. In Southeast Asia, malaria reached 28 million cases by 2010. Artemisinin is a secondary matebolite produced by Artemisia annua L. which is used as a first choice drug in form of ACTs (artemisinin-based combined therapies) to treat malaria as recommended by World Health Organization (WHO). The key gene in the biosynthesis of artemisinin, amorpha-4,11-diene synthase (ads), has become the main research object to enhance production of artemisinin by increasing expression of this gene. The efficiency of transient expression can be elevated by addition of p19 gene. The purpose of this study is to increase efficiency of transient-based transformation through the addition of p19 gene. The p19 gene was inserted at pCAMBIA 1303 and pCAMBIA 1303-ads. The presence of p19 gene was confirmed through analysis of PCR, restriction, and sequencing. The recombinant plasmid obtained were transformed into Agrobacterium tumefaciens AGL 1. Transformation of A. annua was mediated with Agrobacterium harbouring PCAMBIA 1303¬p19-ads, co-transformation was performed using pCAMBIA 1303-ads together wih PCAMBIA 1303-p19. and transformation with pCAMBIA 1303 and pCAMBIA 1303-ads as a control. GUS transient analysis showed that the transformation efficiency of pCAMBIA 1303- ads-p19 (70.89 ± 23.61 %) was higher than pCAMBIA 1303-ads (48.59 ± 38.90 %). The co- transformation efficiency pCAMI3IA 1303-ads and pCAMBIA 1303-p19 (72.49 + 22.16 %) was higher than pCAMBIA 1303-ads-p19 and pCAMBIA 1303-ads. the transformation efficiency of pCAMBIA 1303-p/9 (24.99 ± 17.65 %) was higher than pCAMBIA 1303 (13.70 ± 739 %). Histochemical analysis of leaves culture of A. annua L. showed that the addition of p19 gene gave an influence on the transformation. The increasing of GUS expression was assumed that the ads expression would also enhance leading to the enhance of artemisinin production.
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