Based on the reports from World Health Organization, the malaria disease threatens people’s lives in 106 endemic countries and territories, particularly those countries in Africa. In 2010, approximately 3.3 billion people were at risk of malaria and 655, 000 deaths (particularly children under five years old and pregnant women) were estimated. Plasmodium falciparum that is a resistant parasite to most antimalarial drug causes nearly 80% of casualties. Currently, the most effective treatment of falciparum-caused malaria is the use of artemisinin and the artemisinin-based combination therapies (ACT).
Although several genes (e.g. ADS and CYP71AV1) have been cloned from A. annua and biochemically shown the involvement into the biosynthesis of artemisinin precursors, to date, whether these genes are genetically involved in the biosynthesis of artemisinin has not been investigated. This is because genetic evidence of these gene functions In Planta is lacking. Our goal underlying this specific proposal is to use transgene to demonstrate the functions of CYP71AV1 gene and to increase the production of artemisininin plants. We have cloned CYP71AV1 gene from A. annua and constructed binary vectors for genetic transformation. The experimental procedures of transformation include activation of Agrobacterium, pre-culture of leaf explants,infection of explants by activated Agrobacterium, cultivation of infected explants, regeneration and selection of transgenic plants, identification of transgenic plants, RT-PCR, southern blotting analysis and metabolic analysis of artemisinin and its precursors. We are expecting to obtain multiple transgenic plants with high levels of artemisinic acids and likely artemisinin.