Constant and variable regions in DNA mini-circles from Trypanosoma cruzi and Trypanosoma rangeli: Application to species and stock differentiation

Frasch, Alberto C.C.; Goijman, Silvia G.; Cazzulo, Juan J.; Stoppani, A. O. M.

doi:10.1016/0166-6851(81)90015-3

Cited by 36 publications

(7 citation statements)

References 13 publications

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“…We also found that sequences with aries and that may be analogous to conserved regions found in other kinetoplastida minicircles (24,26,27). There thus appears to be a gradient of sequence variation extending leftward, with sequences conserved in the L. mexicana complex present in the next 170 bp and sequences that have the highest divergence in the leftmost 400 bp (Fig.…”

Section: Discussionmentioning

confidence: 64%

“…Analogous Within a given sequence class, the minicircle sequences are identical (24,25). Sequence analysis of different classes of minicircles in Leishmania tarentole (7) and T. brucei (26) has revealed that minicircles of different classes share =150 bp of homology but are widely divergent throughout the rest of their length.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, the cross-reaction with strains of Old World cutaneous and visceral leishmaniasis can be localized to a maximum of 230 bp from the Dra I site in the minicircle. This may be analogous to the conserved region of minicircle DNA described in other kinetoplastida organisms (7,24) and may be important to minicircle function. The most striking result is presented in Fig.…”

Section: Methodsmentioning

confidence: 99%

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Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Rogers¹,

Wirth²

1987

Proc. Natl. Acad. Sci. U.S.A.

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Previous work has shown that the kinetoplast minicircle DNA of Leishmania species exhibits species-specific sequence divergence and this observation has led to the development of a DNA probe-based diagnostic test for leishmaniasis. In the work reported here, we demonstrate that the minicircle is composed of three types of DNA sequences with differing specificities reflecting different rates of DNA sequence change. A library of cloned fragments of kinetoplast DNA (kDNA) from Leishmania mexicana amazonensis was prepared and the cloned subfragments were found to contain DNA sequences with different taxonomic specificities based on hybridization analysis with various species of Leishmania. Four groups of subfragments were found, those that hybridized with a large number of Leishmania sp. as well as sequences unique to the species, subspecies, or isolate. Analysis of nested deletions of a single, full-length minicircle demonstrates that these different taxonomic specificities are contained within a single minicircle. This implies that different regions of a single minicircle have DNA sequences that diverge at different rates. These sequences represent potentially valuable tools in diagnostic, epidemiologic, and ecological studies of leishmaniasis and provide the basis for a model of kDNA sequence evolution.

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Rogers¹,

Wirth²

1987

Proc. Natl. Acad. Sci. U.S.A.

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“…Previous work on sequence relationships among kDNA minicircles from different genera has shown that a kDNA network consists of a population of minicircles of different sequence and that the degree of sequence heterogeneity varies among different genera [6-81. The occurrence of sequence heterogeneity within the major component of kDNA has been exploited as a genotypic marker to compare strains and clones of T cruzi [3,4,14], T brucei [7,15,16], Crithidia [5,19], cutaneous isolates of Leishmania [ 11,121, and other trypanosomatids [5,14].…”

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confidence: 99%

Characterization of isolates and clones of leishmania by analysis of kinetoplast DNA

Spithill

Grumont

Mitchell

1984

J of Cellular Biochemistry

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The genetic characterization of pathogenic isolates of Leishmania was attempted by analysis of the molecular properties of kinetoplast DNA (kDNA) minicircles. Unit minicircle size is not conserved during speciation of Leishmania since the minicircles of strains and clones of L t major are smaller (700 bp) than those found in certain strains of L mexicana ssp (820 bp), L donovani (850 bp) or L t tropica (900 bp). Schizodeme analysis of minicircles reveals a high degree of sequence divergence in kDNA of Leishmania with the degree of microheterogeneity varying between species. This sequence divergence allows the discrimination of species, strains, and clones of Leishmania into schizodemes . Southern blot hybridization experiments reveal that at high stringency overall minicircle sequence homology is conserved among clones and strains of one species (L t major) but not between different species. This property of minicircle DNA permits the use of kDNA probes as a species-specific diagnostic test for the identification of unknown Leishmania isolates. The properties of kDNA from an L t tropica strain LRC- L32 (a " recidiva " organism) are so diverged from those of L t major strains as to support the classification [22, 23] of L t tropica and L t major as separate species of Leishmania rather than subspecies of L tropica.

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“…Thus, a rapid identification method for T. cruzz isolates and clones would be a valuable tool not only for field studies but also for laboratory work. Among the most powerful methods used are isoenzyme analysis [4,5] and restriction enzyme patterns from the mitochondrially located kinetoplast DNA (kDNA) [3,6], both of which, however, are time-consuming.…”

Section: Introductionmentioning

confidence: 99%

Rapid identification of Trypanosoma cruzi isolates by ‘dot‐spot’ hybridization

Sánchez¹,

Madrid²,

Engel

et al. 1984

FEBS Letters

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The presence of isolate-specific Trypanosoma cruz~ minicircles has been shown in the kinetoplast DNA of this parasite. This led to the rapid identification of isolates and clones of trypanosomes by means of 'dot-spot' hybridizations with molecularly cloned minicircle probes. Unexpectedly, whole kDNAs were also suitable as probes for this purpose, provided that filters were washed under stringent conditions. This was attributed to the presence of the above-mentioned isolate-specific minicircle sequences. The fact that parasites could be directly spotted onto nitrocellulose filters simplified the rapid routine screening of a large number of samples.

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Constant and variable regions in DNA mini-circles from Trypanosoma cruzi and Trypanosoma rangeli: Application to species and stock differentiation

Cited by 36 publications

References 13 publications

Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Characterization of isolates and clones of leishmania by analysis of kinetoplast DNA

Rapid identification of Trypanosoma cruzi isolates by ‘dot‐spot’ hybridization

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