ABSTRACT

Leishmaniasis is caused by an obligate intracellular protozoa belonging to Leishmania genus and listed among major tropical diseases by WHO. Because of the high costs, toxicity, and adverse effects of routinely used compounds in the treatment, alternative treatment and vaccine studies are underway. An effective vaccine has not been developed to date. In this study, we aimed to clone one of the most promising DNA vaccine candidates: the homolog-activated C kinase (LACK) gene of Leishmania infantum.

L. infantum genomic DNA was isolated from promastigote culture. The LACK gene was placed into plasmid pJET1.2. Then, recombinant plasmids were transformed into competent cells. The presence of recombinant plasmids was determined by PCR screening. Cloning was confirmed by PCR, restriction enzyme assays, and finally, DNA sequence analysis, after making miniprep from positive colonies.

After performing PCR with LACK-gene specific primers, 939-bp PCR products were observed. Recombinant plasmids, which were transformed into competent Escherichia coli cells, were verified by PCR screening. It was verified by PCR that the recombinant plasmid contained the LACK gene. DNA sequence analysis was performed to obtain the DNA sequence.

One of the most promising DNA vaccine candidates against leishmaniasis, the LACK gene, was cloned in this study.

Keywords: Leishmaniasis, Leishmania infantum, LACK gene, DNA vaccine, cloning