Lewis, E., Hunter, S. J., Tetley, L., Nunes, C. P., Bazzicalupo, P. and Devaney, E. (1999) cut-1-like genes are present in the filarial nematodes, Brugia pahangi and Brugia malayi, and, as in other nematodes, code for components of the cuticle. Molecular and Biochemical Parasitology, 101(1-2), pp. 173-83. (doi: 10.1016/S0166-6851(99)00070-5) (PMID:10413052)
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Abstract
A fragment of a cut-1 like gene from the filarial nematode Brugia pahangi (designated Bp-cut-1) was isolated by PCR from genomic DNA. The sequence was used to design primers for use in RT-PCR and resulted in the isolation of a cDNA fragment from larvae in the process of the L3-L4 moult. Screening of a B. malayi genomic library identified a single clone, Bm-cut-1. Using primers designed from the Brugia sequences, semi-quantitative RT-PCR was carried out on 11 different life cycle stages chosen to cover periods around the moult and inter-moult periods. This analysis demonstrated that the cut-1 mRNA was most abundant preceding the moult, consistent with its function as a cuticular protein. Immuno-gold electron microscopy using an affinity purified antiserum raised to the highly conserved region of Ascaris CUT-1 confirmed that the protein was restricted to a tight band in the median layer of the cuticle. Despite the fact that no transcripts could be detected in mature adult worms by RT-PCR, immuno-gold microscopy revealed staining of the microfilarial cuticle within the uterus of the adult female worm, suggesting that other cut-1-like genes are present in Brugia.
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Tetley, Dr Laurence and Devaney, Professor Eileen |
Authors: | Lewis, E., Hunter, S. J., Tetley, L., Nunes, C. P., Bazzicalupo, P., and Devaney, E. |
College/School: | College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine |
Journal Name: | Molecular and Biochemical Parasitology |
Publisher: | Elsevier BV |
ISSN: | 0166-6851 |
ISSN (Online): | 1872-9428 |
Published Online: | 20 February 2003 |
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