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Isolation and characterization of a fatty acid- and retinoid-binding protein from the cereal cyst nematode Heterodera avenae.
- Source :
-
Experimental parasitology [Exp Parasitol] 2016 Aug; Vol. 167, pp. 94-102. Date of Electronic Publication: 2016 May 27. - Publication Year :
- 2016
-
Abstract
- A gene encoding fatty acid- and retinoid-binding protein was isolated from the cereal cyst nematode Heterodera avenae and the biochemical function of the protein that it encodes was analysed. The full-length cDNA of the Ha-far-1 gene is 827 bp long and includes a 22- nucleotide trans-spliced leader sequence (SL1) at its 5-end. The genomic clone of Ha-far-1 consists of eight exons separated by seven introns, which range in size from 48 to 186 bp. The Ha-far-1 cDNA contains an open reading frame encoding a 191 amino acid protein, with a predicted secretory signal peptide. Sequence analysis showed that Ha-FAR-1 has highest similarity to the Gp-FAR-1 protein from the potato cyst nematode, Globodera pallida and that the protein was grouped with all homologues from other plant-parasitic nematodes in a phylogenetic analysis. Fluorescence-based ligand binding analysis confirmed that the recombinant Ha-FAR-1 protein was able to bind fatty acids and retinol. Spatial and temporal expression assays showed that the transcripts of Ha-far-1 accumulated mainly in the hypodermis and that the gene is most highly expressed in third-stage juveniles of H. avenae. Fluorescence immunolocalization showed that the Ha-FAR-1 protein was present on the surface of the infective second-stage juveniles of H. avenae. Nematodes treated with dsRNA corresponding to Ha-far-1 showed significantly reduced reproduction compared to nematodes exposed to dsRNA from a non-endogenous gene, suggesting that Ha-far-1 may be an effective target gene for control of H. avenae using an RNAi strategy.<br /> (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Blotting, Western
Cloning, Molecular
DNA, Complementary chemistry
DNA, Helminth isolation & purification
Fatty Acid-Binding Proteins chemistry
Fatty Acid-Binding Proteins genetics
Female
Gene Expression Regulation, Developmental
Gene Silencing
Helminth Proteins chemistry
Helminth Proteins genetics
In Situ Hybridization
Ligands
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Retinol-Binding Proteins chemistry
Retinol-Binding Proteins genetics
Sequence Alignment
Transcription, Genetic
Triticum parasitology
Tylenchoidea genetics
Fatty Acid-Binding Proteins isolation & purification
Helminth Proteins isolation & purification
Retinol-Binding Proteins isolation & purification
Tylenchoidea chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1090-2449
- Volume :
- 167
- Database :
- MEDLINE
- Journal :
- Experimental parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 27240755
- Full Text :
- https://doi.org/10.1016/j.exppara.2016.05.009