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Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum

Investigation article published in International journal for parasitology

July 1st, 2007

Here we present the identification and cloning of the NcBSR4 gene, the putative Neospora caninum orthologue to the Toxoplasma gondii TgBSR4 gene. To isolate NcBSR4, genome walking PCR was performed on N. caninum genomic DNA using the expressed sequence tag NcEST3c28h02.y1 sequence, which shares a 44% identity with the TgBSR4 gene, as a framework. Nucleotide sequencing of amplified DNA fragments revealed a single uninterrupted 1227 bp open reading frame that encodes a protein of 408 amino acids with 66% similarity to the TgBSR4 antigen. A putative 39-residue signal peptide was found at the NH2-terminus, followed by a hydrophilic region. At the COOH-terminus, a potential site for a glycosylphosphatidylinositol anchor was identified at amino acid 379. A polyclonal serum against recombinant NcBSR4 protein was raised in rabbits, and immunolabelling demonstrated stage-specific expression of the NcBSR4 antigen in N. caninum bradyzoites produced in vitro and in vivo. Furthermore, RT-PCR analysis showed a slight increase of NcBSR4 transcripts in bradyzoites generated during in vitro tachyzoite-to-bradyzoite stage-conversion, suggesting that this gene is specifically expressed at the bradyzoite stage and that its transcription relies on the switch to this stage




Risco-Castillo V., Fernandez-Garcia A., Zaballos A., Aguado-Martinez A., Hemphill A., Rodriguez-Bertos A., Alvarez-Garcia G. and Ortega-Mora LM.




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Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum

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Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum



Participants:

Universidad Autónoma de MadridUniversidad Autónoma de Madrid (UAM).

Consejo Superior de Investigaciones CientíficasDpto. de Inmunología y Oncología. Centro Nacional de Biotecnología (CNB). Consejo Superior de Investigaciones Científicas (CSIC).

Universität BernInstitute of Parasitology. Vetsuisse-Fakultät. Universität Bern (UB).

Universidad ComplutenseServicio de Identificación y Caracterización Microbiana (ICM). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

Universidad ComplutenseSalud Veterinaria y Zoonosis (SALUVET). Departamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).







FACTOR YEAR Q
3.392 2007

NLMID: 314024

PMID: 17376452

ISSN: 0020-7519



TITLE: Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum


JOURNAL: Int J Parasitol


NUMERACIÓN: 37(8-9):887-96


AÑO: 2007


PUBLISHER: Oxford, New York, Pergamon Press


AUTHORS: Risco-Castillo V., Fernandez-Garcia A., Zaballos A., Aguado-Martinez A., Hemphill A., Rodriguez-Bertos A., Alvarez-Garcia G. and Ortega-Mora LM.


Antonio Manuel Rodríguez Bertos

DOI: https://doi.org/10.1016/j.ijpara.2007.02.003


CITE THIS PUBLICATION:

Risco-Castillo V., Fernandez-Garcia A., Zaballos A., Aguado-Martinez A., Hemphill A., Rodriguez-Bertos A., Alvarez-Garcia G. and Ortega-Mora LM. Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum. International journal for parasitology. 37(8-9):887-96. 2007. (A). ISSN: 0020-7519. DOI: 10.1016/j.ijpara.2007.02.003


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