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

Risco-Castillo, V; Fernández-García, A; Zaballos, A; Aguado-Martínez, A; Hemphill, A; Rodríguez-Bertos, A; Alvarez-García, G; Ortega-Mora, L M (2007). Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum. International journal for parasitology, 37(8-9), pp. 887-96. Kidlington, UK: Elsevier 10.1016/j.ijpara.2007.02.003

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology

UniBE Contributor:

Hemphill, Andrew

ISSN:

0020-7519

Publisher:

Elsevier

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:54

Last Modified:

05 Dec 2022 14:16

Publisher DOI:

10.1016/j.ijpara.2007.02.003

PubMed ID:

17376452

Web of Science ID:

000248068800008

URI:

https://boris.unibe.ch/id/eprint/22737 (FactScience: 36374)

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