The β-Tubulin Gene Region as a Molecular Marker to Distinguish Leishmania Parasites

Mendoza-León, Alexis; Luis, Luis; Martı́nez, Clara

doi:10.1385/1-59259-238-4:061

Cited by 7 publications

(7 citation statements)

References 17 publications

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“…Two markers, previously reported as fit to evaluate genetic diversity in different organisms, were used in T. maculata . First, the mt Cyt b gene apply to evaluate population genetic structure in different triatomine species ( 28 , 29 ), and second, the β-tubulin gene region, a marker employ to study genetic variability in Leishmania subgenera ( 30 , 31 ).…”

Section: Methodsmentioning

confidence: 99%

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat

García-Alzate

Lozano-Arias

Reyes-Lugo

et al. 2014

Front. Public Health

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Triatoma maculata is a wild vector of Trypanosoma cruzi, the causative agent of Chagas disease; its incursion in the domestic habitat is scant. In order to establish the possible domestic habitat of T. maculata, we evaluated wing variability and polymorphism of genotypic markers in subpopulations of T. maculata that live in different habitats in Venezuela. As markers, we used the mtCyt b gene, previously apply to evaluate population genetic structure in triatomine species, and the β-tubulin gene region, a marker employed to study genetic variability in Leishmania subgenera. Adults of T. maculata were captured in the period 2012–2013 at domestic, peridomestic (PD), and wild areas of towns in the Venezuelan states of Anzoátegui, Bolívar, Portuguesa, Monagas, Nueva Esparta, and Sucre. The phenotypic analysis was conducted through the determination of the isometric size and conformation of the left wing of each insect (492 individuals), using the MorphoJ program. Results reveal that insects of the domestic habitat showed significant reductions in wing size and variations in anatomical characteristics associated with flying, in relation to the PD and wild habitats. The largest variability was found in Anzoátegui and Monagas. The genotypic variability was assessed by in silico sequence comparison of the molecular markers and PCR-RFLP assays, demonstrating a marked polymorphism for the markers in insects of the domestic habitat in comparison with the other habitats. The highest polymorphism was found for the β-tubulin marker with enzymes BamHI and KpnI. Additionally, the infection rate by T. cruzi was higher in Monagas and Sucre (26.8 and 37.0%, respectively), while in domestic habitats the infestation rate was highest in Anzoátegui (22.3%). Results suggest domestic habitat colonization by T. maculata that in epidemiological terms, coupled with the presence in this habitat of nymphs of the vector, represents a high risk of transmission of Chagas disease.

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Section: Methodsmentioning

confidence: 99%

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat

García-Alzate

Lozano-Arias

Reyes-Lugo

et al. 2014

Front. Public Health

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“…Even though many investigations indicate that genetic variation in parasitic nematodes is an issue of considerable practical and theoretical significance, either from a morphological (Mendoza-Leon, Luis & Martinez, 2001), biological (Watkins & Fernando, 1984), therapeutic (Hejmadi et al 2000) and immunological (Goyal & Wakelin, 1993) point of view, the extent to which genetically determined variation affects the host-parasite interaction is still poorly understood (Wakelin, Farias & Bradley, 2002). Much more work is needed to elucidate the functional implications of the genetic variability in cysteine protease genes from H. contortus measured in this and previous studies.…”

Section: Discussionmentioning

confidence: 83%

Genetic variability in cysteine protease genes ofHaemonchus contortus

et al. 2004

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To increase the existent genetic variability in cysteine proteases, a polymorphism study was performed in Haemonchus contortus by comparing 2 different strains of the parasite: North American (NA) and Spanish (SP) strains. For this purpose, the polymorphism of 5 previously reported genes (AC-1, AC-3, AC-4, AC-5 and GCP-7) were analysed by PCR-SSCP and sequencing procedures. Based on the SSCP results, a total of 20 different alleles were identified for the 5 loci assessed. Except locus AC-5, all the loci were polymorphic. Loci AC-1, AC-3, AC-4 and GCP-7 showed 5, 8, 2 and 4 alleles, respectively. The allelic frequencies ranged from 0.0070 to 0.8560 and were significantly different between strains. In addition, nucleotide diversity analyses showed a significant variation within and between strains. The variations in the nucleotide sequence of the different alleles were translated in some cases into changes in the amino acid sequence. Evidence of genetic variability in cysteine proteases from two different strains of H. contortus for the same set of genes had not been previously reported.

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“…For all analyses, promastigotes were grown at room temperature in Schneider's Drosophila medium (Gibco) supplemented with 10% heat-inactivated fetal calf serum and 100 g/ml kanamycin [Mendoza-León et al, 2002]. The parasite pellet (3 !…”

Section: Methodsmentioning

confidence: 99%

The 6-Phosphogluconate Dehydrogenase of Leishmania (Leishmania) mexicana: Gene Characterization and Protein Structure Prediction

González

Pérez²,

Serrano³

et al. 2010

Microb Physiol

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6-Phosphogluconate dehydrogenase (6PGDH) is a key enzyme of the oxidative branch involved in the generation of NADPH and ribulose 5-phosphate. In the present work, we describe the cloning, sequencing and characterization of a 6PGDH gene from Leishmania (Leishmania) mexicana. The gene encodes a polypeptide chain of 479 amino acid residues with a predicted molecular mass of 52 kDa and a pI of 5.77. The recombinant protein possesses a dimeric quaternary structure and displays kinetic parameter values intermediate between those reported for Trypanosoma brucei and T. cruzi with apparent K_m values of 6.93 and 5.2 µM for 6PG and NADP⁺, respectively. The three-dimensional structure of the enzymes of Leishmania and T. cruzi were modelled from their amino acid sequence using the crystal structure of the enzyme of T. brucei as template. The amino acid residues located in the 6PGDH C-terminal region, which are known to participate in the salt bridges maintaining the protein dimeric structure, differed significantly among the enzymes of Leishmania, T. cruzi, and T. brucei. Our results strongly suggest that 6PGDH can be selected as a potential target for the development of new therapeutic drugs in order to improve existing chemotherapeutic treatments against these parasites.

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The β-Tubulin Gene Region as a Molecular Marker to Distinguish Leishmania Parasites

Cited by 7 publications

References 17 publications

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat

Genetic variability in cysteine protease genes ofHaemonchus contortus

The 6-Phosphogluconate Dehydrogenase of Leishmania (Leishmania) mexicana: Gene Characterization and Protein Structure Prediction

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