The Hamster Model for Identification of Specific Antigens of <i>Taenia solium</i> Tapeworms

Ochoa‐Sánchez, Alicia; Jiménez, Lucı́a; Landa, Abraham

doi:10.1155/2011/504959

Cited by 6 publications

(3 citation statements)

References 31 publications

(44 reference statements)

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“…For western blot analysis, 1 μ g/mm of rTsTrx-1, rTrx- E. coli , and human T-cell recombinant Trx (rTrx-human, Sigma-Aldrich, St. Louis. MO) and likewise 5 μ g/mm of parasites crude extract and T. solium cysticerci excretion-secretion antigens (E/S Ag), prepared as described in [ 14 ], were separated by 15% SDS-PAGE and transferred onto a nitrocellulose membrane (Amersham Biosciences, Sweden). The membrane was blocked with 1% BSA in PBS containing 0.05% Tween 20 buffer and incubated for 2 h at room temperature with the anti-rTsTrx-1 serum (dilution 1 : 100).…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Thioredoxin-1 Gene fromTaenia soliumand Its Encoding Product

Jiménez

Rodríguez-Lima

Ochoa‐Sánchez

et al. 2015

BioMed Research International

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Taenia solium thioredoxin-1 gene (TsTrx-1) has a length of 771 bp with three exons and two introns. The core promoter gene presents two putative stress transcription factor binding sites, one putative TATA box, and a transcription start site (TSS). TsTrx-1 mRNA is expressed higher in larvae than in adult. This gene encodes a protein of 107 amino acids that presents the Trx active site (CGPC), the classical secondary structure of the thioredoxin fold, and the highest degree of identity with the Echinococcus granulosus Trx. A recombinant TsTrx-1 (rTsTrx-1) was produced in Escherichia coli with redox activity. Optimal activity for rTsTrx-1 was at pH 6.5 in the range of 15 to 25°C. The enzyme conserved activity for 3 h and lost it in 24 h at 37°C. rTsTrx-1 lost 50% activity after 1 h and lost activity completely in 24 h at temperatures higher than 55°C. Best storage temperature for rTsTrx-1 was at −70°C. It was inhibited by high concentrations of H2O2 and methylglyoxal (MG), but it was inhibited neither by NaCl nor by anti-rTsTrx-1 rabbit antibodies that strongly recognized a ~12 kDa band in extracts from several parasites. These TsTrx-1 properties open the opportunity to study its role in relationship T. solium-hosts.

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Section: Methodsmentioning

confidence: 99%

Characterization of a Thioredoxin-1 Gene fromTaenia soliumand Its Encoding Product

Jiménez

Rodríguez-Lima

Ochoa‐Sánchez

et al. 2015

BioMed Research International

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“…Some T. crassiceps cysticerci were used for confocal microscopy. The T. solium adults were obtained from small intestine of experimental infected hamsters as previously reported [15].…”

Section: Biological Materialsmentioning

confidence: 99%

Taenia soliumTAF6 and TAF9 binds to a putative Downstream Promoter Element present inTsTBP1gene core promoter

Rodríguez-Lima

García-Gutiérrez

Jiménez

et al. 2017

Preprint

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Edificio A 2do piso. Ciudad Universitaria. CDMX 04510, México Phone: (52 55) 56-23-23-57, Fax: (52 55) 56-23-23-82, Email: landap@unam.mx 2 Abstract.We have cloned and characterized the gene encoding to Taenia solium TATA binding protein 1 (TsTBP1). It spans 1481 bp and its coding region is interrupted by four introns that possess the consensus donor/acceptor sequences. It produces a protein of 238 amino acids residues, which presented all the classical motives of the TBP1. On the core promoter region we identified putative binding sites for NF1, AP-1, YY1, TAF1/TAF2 and TAF6/TAF9, and the TSS that corresponds to an A +1 . Southern and Northern blot analysis showed that TsTBP1 is encoded by a single gene, which produce a messenger of about 1.1 kbp with a higher differential expression in adult than the larval stage. Moreover, two putative TATA boxes were identified at -97 and -69 bp (relative to the TSS), likewise a Downstream Promoter Element (DPE) was located at +27 to +31 bp. EMSA experiments did not show any component of cysticerci nuclear extracts bound to the putative TATA-box elements; in contrast TAF6 and TAF9 bind to the DPE, showing that TsTBP1 is a TATAless gen. On the other hand, TAF6 and TAF9 were localized on the nucleus from cells of T. crassiceps cysticerci bladder walls. By using the amino acid sequences of T. solium TAF6 (TsTAF6) and TAF9 (TsTAF9), we constructed a molecular model showing interaction between DPE-TsTAF6 and TsTAF6-TsTAF9. Finally, an in silico analysis of the core promoters of Taeniidae family genes showed that TATA-box and DPE are homologous to the mammalian elements, but not so the Inr. The low identity between TAF9 from cestodes and mammals open the possibility to use it as target to interrupt or modulate the transcription of TATA-box less genes in cestodes as a novel therapeutic strategy. 3 Author summary. Neurocysticercosis still being a health problem in developing countries. Mexico reports a prevalence of 2.5% in the total patients from National Institute of Neurology and Neurosurgery. Several efforts have been made in different fields to study Taenia solium, and although the Genome Project has been published and released the genomic sequence of this parasite; the transcriptional mechanisms this organism remains unexplored. We isolated and characterized Taenia solium TATA-Binding Protein 1 (TsTBP1) gene and TBP-associated factor 6 and 9 cDNAs (TsTAF6 and TsTAF9, respectively). Our principal findings are: 1.-Identification of cis and trans elements in the core promoter of TsTBP1 gene. 2.-The TsTBP1 gene is a TATA-less promoter. 3.-Cloning of cDNAs to TsTAF6 and TsTAF9, 4.-Identification of a DPE in the core promoter of TsTBP1 gene, which interacts with TAF6 and TAF9, 5.-Construction of a molecular model that shows the interaction between DPE, TAF6, and TAF9; and 6.-A proposal of consensus sequences for TATAbox, Inr and DPE presented on Taeniidae family core promoters.4

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“…39 Existen varias técnicas diagnósticas, pero ninguna de ellas se ha podido implementar para llevar a cabo un diagnóstico masivo, ya que debe tomarse en cuenta que las zonas de mayor prevalencia de la infección son de escasos recursos económicos y el costo de estas técnicas es elevado. Las técnicas que se han desarrollado son: la detección de coproantígenos por un ensayo inmunoenzimático (ELISA), en el cual son detectadas moléculas específicas del cestodo en muestras fecales; 59,60,61 la reacción en cadena de la polimerasa (PCR) para amplificar genes mitocondriales del ADN proveniente de huevos; 62, 32 63 e inmunoelectrotransferencia (IET) para detectar moléculas especificas del parásito, como los antígenos de excreción-secreción; 64,65,66 la búsqueda de moléculas especificas de este estadio aún continúa. Actualmente se intenta usar proteínas recombinantes.…”

Section: Teniosisunclassified

Taenia solium: anticuerpos como inhibidores de la actividad de la Triosafosfato isomerasa (TPI)

Ayala¹

2013

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The Hamster Model for Identification of Specific Antigens of Taenia solium Tapeworms

Cited by 6 publications

References 31 publications

Characterization of a Thioredoxin-1 Gene fromTaenia soliumand Its Encoding Product

Characterization of a Thioredoxin-1 Gene fromTaenia soliumand Its Encoding Product

Taenia soliumTAF6 and TAF9 binds to a putative Downstream Promoter Element present inTsTBP1gene core promoter

Taenia solium: anticuerpos como inhibidores de la actividad de la Triosafosfato isomerasa (TPI)

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