Journal of Biotechnology & Biomaterials
Open Access

Abstract

Malaria is still remaining 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 as 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 its expression. The efficiency of transient expression can be elevated by addition of p19 gene. The purpose of this study was 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 plasmids obtained were transformed into Agrobacterium tumefaciens AGL1. 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 pCAMBIA 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-p19 (24.99±17.65%) was higher than pCAMBIA 1303 (13.70±7.39%). Histochemical analysis of leaves culture of A. annua L. showed that the addition of p19 gene could enhance the transformation efficiency of amorphadienesunthase in Artemisia annua . The increasing of GUS expression was assumed that the ads expression would also enhance leading to enhanced artemisinin production.

Biography

Elfahmi has completed his PhD from University of Groningen, the Netherlands, 2006. He is the Head of Graduate Program at the School of Pharmacy, ITB, Indonesia, and as consultant at Indonesian FDA (BPOM). Beside as a lecturer (an associate professor), he is also very active in research in the field of herbal medicines. He has published more than 20 papers in reputed journals and has been serving as reviewers in some international and national journals. He has presented his research result in many national and international seminars.