Molecular Cloning and Expression of the Leishmania infantum KMP-11 Gene

Abstract

Background: Visceral leishmaniasis (Kala-azar) is one of the most serious tropical diseases, and it can lead to death. The kinetoplastid membrane protein-11 (KMP-11) is highly conserved in all stages of the Leishmania life cycle. Objectives: In the present study, the KMP-11 gene was extracted from Leishmania infantum and then, cloned and expressed in an expression vector.The main objective of this study was to provide an introduction to DNA vaccine production against visceral leishmaniasis. Materials and Methods: DNA was extracted from L. infantum (MHOM/TN/80/IPI1), and amplified by PCR and a specific primer. Afterwards the purified PCR products were successfully ligated into the pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as an expression vector and the KMP-11 gene subcloned into this plasmid. The pTKMP-11 plasmid was digested by restriction enzymes to confirm cloning of this gene in the pTZ57R/T plasmid. In the last step, the subcloned gene was expressed in a eukaryotic cell. Results: The KMP-11 gene was successfully subcloned in pcDNA3 as a eukaryotic expression vector. Recombinant KMP-11 protein was confirmed by the reverse transcriptase polymerase chain reaction (RT-PCR) method. Conclusions: The results of the present study will increase our knowledge about molecular cloning and expression of the L. infantum KMP-11 gene, and this may be used as an effective target for controlling visceral leishmaniasis.