Molecuar Analysis of Peroxidoxin Genes from Leishmania Aethiopica

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Date

2004-06

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Addis Ababa Universty

Abstract

Cutaneous leishmaniasis in Ethiopia is primarily caused by Leishmania aethiopica and rarely by L. tropica and L. major. There is no rapid differential diagnostic method for these species. The intracellular survival mechanism of Leishmania parasites is not well understood although one of the mechanisms may be production of antioxidant enzymes such as peroxidoxins. Peroxidoxin genes have been isolated from various Leishmania species but not from L. aethiopica. Our objective was to identify and characterize peroxidoxin genes from L. aethiopica. In this study, we identified two peroxidoxin genes (pxnl and Pxn2) from L. aethiopica by PCR. Sequence analysis of the two genes showed that there is a high nucleotide sequence homology between L. aetlziopica and other Leishmania species. Southern blot hybridization analysis showed peroxidoxins of L. aetlziopica genes exist as multigene family. RT-PCR demonstrated that pxnl is predominantly expressed in amastigotes and stationary phase promastigotes, suggesting its importance for infectivity and intracellular survival. Pxn2 is constitutively expressed in the different stages of the parasite. Northern blot analysis and RT-PCR showed that the overall expression of all peroxidoxin genes in L. aethiopica is higher in the amastigotes than in the promastigotes, suggesting that peroxidoxins are impoliant for intracellular survival. Peroxidoxin genes appear to be important virulence factors and thus may be potential targets for drug development against leishmaniasis.The genes can be explored further as potential candidate vaccines and molecular diagnostic tools. We suggest that further research be conducted on peroxidoxins to evaluate their potential as diagnostics, vaccine candidates and drug targets. Key words: Intracellular survival, L. aethiopica, peroxidoxin genes

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Keywords

Intracellular survival, L. aethiopica, Peroxidoxin genes

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