Kloning gen CYP71AV asal Artemisia annua di dalam Escherichia coli DH-5α

Abstract

Infection of malaria caused 1.7-2.5 million people die each year. Artemisia annua contains artemisinin compounds effective to eradicate malaria types that are resistant to quinine and chloroquine. CYP71AV is a gene that has critical role in the biosynthesis of artemisinin by catalyzing the oxidation of intermediate products Amorpha-4,11-diene, which are highly produced in the biosynthesis of artemisinin. This study aimed to clone the CYP71AV gene through recombinant DNA technology. CYP71AV gene from total cDNA was amplified using specific primers. CYP71AV gene was inserted into pGEM-T Easy plasmid. The recombinant plasmid containing CYP71AV gene introduced into Escherichia coli strain DH5-α. CYP71AV gene sequencing produce approximately 1571 base pairs encoding 523 amino acids. Local alignment analysis of nucleotides with CYP71AV of several species listed in GeneBank showed that the cDNA CYP71AV have 98% similarity with complete cds A.annua (JQ254992.1). Domain analysis showed that the CYP71AV gene has three conserved domain