Molecular Characterization of Superoxide Dismutase Gene From Leishmania Aethiopica
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Date
2003-06
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Addis Ababa University
Abstract
This study was perfOlmed with the aim of characterizing the genes that code for
superoxide dismutase in Leishmania aethiopica. It involved three main steps:
specimen collection and parasite isolation, species typing, and molecular
characterization of the SOD genes. A total of 20 suspected cutaneous leishmaniasis
patients were enrolled in the study. Clinical and epidemiological infOlmation was
collected from all the subjects including age, sex, address, duration of the lesion, and
site of lesion. Skin slit was taken from the active lesion(s) and was subjected to ill
vitro parasite culture on NNN medium. Promastigote stage of Leishmania was then
isolated and characterized. Species typing was done on the isolate using isoenzyme
electrophoresis and PCR-RFLP. Total genomic DNA was extracted from the
promastigotes and then the SOD genes were amplified using polymerase chain
reaction. The amplified fragments were then sequenced and the sequence information
was analyzed using vector NTI software.
The study subjects came from different parts of the countLy including Addis Ababa.
The mean age of the patients was 25.6 year with a range of 7 to 65 years. Out of 20
skin slit specimens cultured and processed from suspected cutaneous leishmaniasis
patients enrolled in the study, five (25%) were found to be positive for motile
promastigotes. Isoenzyme electrophoresis and PCR-RFLP results of one isolate
(1093/02) confilmed that the isolate is L. aethiopica. The SODB genes amplified from
promastigotes of L. aethiopica were similar in size as the SODB genes of other
·Leishmania species. Nucleotide sequences of LaethFeSODBI showed 94.9% and 73%
identity with LcFeSODB I and LmFeSODBI respectively. On the other hand,
predicted amino acid sequence comparison indicated that LaethFeSODB 1 had 89.8%
and 90.8% identity with LcFeSODB 1 and LmFeSODB I respectively. The high degree
of amino acid sequence similarity of L. aetlziopica SODB genes with those of other
species has an important implication in the use of new antileishmanial drugs that are
targeting SODs of other Leishmania species to treat cutaneous leishmaniasis in
Ethiopia.
Key words: Leishmallia aethiopica, superoxide dismutase, gene, Ethiopia
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Keywords
Leishmallia aethiopica, superoxide dismutase, gene, Ethiopia