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Leishmania parasites and their kinetoplast DNA (kDNA)

Published online by Cambridge University Press:  19 September 2011

N. N. Massamba ,
M. J. Mutinga  and
B. N. Odero
N. N. Massamba
Affiliation:
The International Centre of Insect Physiology and Ecology (ICIPE), P. O. Box 30772, Nairobi, Kenya
M. J. Mutinga
Affiliation:
The International Centre of Insect Physiology and Ecology (ICIPE), P. O. Box 30772, Nairobi, Kenya
B. N. Odero
Affiliation:
The International Centre of Insect Physiology and Ecology (ICIPE), P. O. Box 30772, Nairobi, Kenya
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Abstract

Restriction endonuclease digestion of genomic DNAs from WHO Leishmania reference strains generates prominent kinetoplast DNA (kDNA) fragments ranging from 0.5–2.0 kb. Following fractionation by agarose gel electrophoresis, these DNA fragments were visualized as distinct bands in gels. Using as parameters the presence or absence of the prominent bands and their size and number, the Leishmania reference strains fell into three distinct genomic groups. These genomic groupings were applied to new Leishmania isolates. DNA analysis, involving restriction endonuclease digestion and Southern blot hybridization with probes generated from the prominent DNA bands of reference strains L. major IC-236 and L. major IC-235, cloned in plasmids, showed significant genetic variation within Leishmania and allowed some distinct isolates to be identified.

Résumé

La digestion aux enzymes de restriction des ADN génomiques obtenus à partir des souches de référence de Leishmania génère des fragments caractéristiques des ADN (ADN kinétoplastique) dont la taille est comprise entre 0.5–2.0 kb. Le fractionnement par électrophorèse sur gel d'agarose de ces fragments d'ADN révèle des bandes proéminentes distinctes. En se servant comme paramètres, la présence ou l'absence de ces bandes, leur taille et nombre, il a été possible de répartir les souches de référence de Leishmania en trois différents groupes génomiques. L'application de ce mode de partage aux nouveaux isolats est rapportée dans cette étude. L'analyse de l'ADN, faisant intervenir la digestion aux enzymes de restriction, l'hybridation à la Southern avec des sondes préparées à partir des fragments caractéristiques de l'ADN des souches de référence L. major IC-236 et L. major IC-235, clonés dans les plasmides a montré une variation génétique significative au sein des Leishmania et quel ques isolats ont été ainsi identifiés.

Keywords

WHO Leishmania reference strainskinetoplast DNA (kDNA)kilobase (kb)DNA probessouthern blot hybridizationSouches de référence de LeishmaniaADN kinétoplastiquekilobase (kb)sondes d'ADNhybridation selon southern
Type
Research Articles
Information
International Journal of Tropical Insect Science , Volume 13 , Issue 3 , June 1992 , pp. 293 - 306
Copyright
Copyright © ICIPE 1992

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References

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