Molecular Characterization Of Superoxide Dismutase Gene From Leishmania Aethiopica

This study was perfOlmed with the aim of characterizing the genes that code forrnsuperoxide dismutase in Leishmania aethiopica. It involved three main steps:rnspecimen collection and parasite isolation, species typing, and molecularrncharacterization of the SOD genes. A total of 20 suspected cutaneous leishmaniasisrnpatients were enrolled in the study. Clinical and epidemiological infOlmation wasrncollected from all the subjects including age, sex, address, duration of the lesion, andrnsite of lesion. Skin slit was taken from the active lesion(s) and was subjected to illrnvitro parasite culture on NNN medium. Promastigote stage of Leishmania was thenrnisolated and characterized. Species typing was done on the isolate using isoenzymernelectrophoresis and PCR-RFLP. Total genomic DNA was extracted from thernpromastigotes and then the SOD genes were amplified using polymerase chainrnreaction. The amplified fragments were then sequenced and the sequence informationrnwas analyzed using vector NTI software.rnThe study subjects came from different parts of the countLy including Addis Ababa.rnThe mean age of the patients was 25.6 year with a range of 7 to 65 years. Out of 20rnskin slit specimens cultured and processed from suspected cutaneous leishmaniasisrnpatients enrolled in the study, five (25%) were found to be positive for motilernpromastigotes. Isoenzyme electrophoresis and PCR-RFLP results of one isolatern(1093/02) confilmed that the isolate is L. aethiopica. The SODB genes amplified fromrnpromastigotes of L. aethiopica were similar in size as the SODB genes of otherrn·Leishmania species. Nucleotide sequences of LaethFeSODBI showed 94.9% and 73%rnidentity with LcFeSODB I and LmFeSODBI respectively. On the other hand,rnpredicted amino acid sequence comparison indicated that LaethFeSODB 1 had 89.8%rnand 90.8% identity with LcFeSODB 1 and LmFeSODB I respectively. The high degreernof amino acid sequence similarity of L. aetlziopica SODB genes with those of otherrnspecies has an important implication in the use of new antileishmanial drugs that arerntargeting SODs of other Leishmania species to treat cutaneous leishmaniasis inrnEthiopia.rnKey words: Leishmallia aethiopica, superoxide dismutase, gene, Ethiopia

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