Detect the presence of LeIF gene in the Leishmania tropica genome and sequence it

Leishmaniasis remains one of the most neglected diseases globally and affects primarily the poor in developing countries. Due to the absence of safe and effective drug, the prevention of infection of this disease is very important as it requires an effective vaccine is not yet available. Leishmania eukaryotic initiation factor (LeIF) protein acts an inhibitor of the Leishmania major growth in murine macrophages and reduces interleukin-4 in lymph nodes. We aimed to detect its presence in Leishmania tropica genome and clone the encoding gene of LeIF antigen of Leishmania tropica into eukaryotic expression plasmid pCI. We used a Syrian strain; LCEB-Syrian Strain 01 of Leishmania tropica promastigotes. Specific primers were designed to amplify the gene by Polymerase Chain Reaction “PCR”. LeIF gene was amplified from both DNA and cDNA after optimization the conditions and then sequenced. LeIF gene was cloned into pCI plasmid. We transformed E. coli Top 10 bacteria by the recombinant plasmid and the cloning was proved by colony PCR and restriction digestion. As a result, LeIF is a part of both DNA genome and RNA of Leishmania tropica LCEB-Syrian Strain 01, and cloned LeIF is a candidate for future studies as a DNA vaccine against leishmaniasis.