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The isolation of differentially expressed cDNA clones from the filarial nematode Brugia pahangi

Hunter, S.J., Martin, S.A., Thompson, F.J., Tetley, L., and Devaney, E. (1999) The isolation of differentially expressed cDNA clones from the filarial nematode Brugia pahangi. Parasitology, 119 (2). pp. 189-198. ISSN 0031-1820 (doi:10.1017/S0031182099004576)

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Abstract

A cDNA library constructed from 3 day post-infective L3 of the filarial nematode Brugia pahangi was screened by differential hybridization with cDNA probes prepared from different life-cycle stages. Five cDNA clones hybridizing selectively to the mosquito-derived L3 probe were isolated and characterized. Northern blot analysis of 4 of the clones confirmed that each was most highly expressed in the mosquito-derived L3. The expression of each mRNA during parasite development in the mosquito vector was investigated using RT-PCR, and all were shown to be abundant in the immature L3. Four of the 5 cDNAs cloned coded for structural proteins: 2 cuticular collagens, and the muscle proteins tropomyosin and troponin. Further studies on troponin using an antiserum raised to the recombinant protein demonstrated that the protein, unlike the mRNA, was present in all life-cycle stages examined, while immunogold labelling demonstrated that it was localized to the muscle blocks.

Item Type:Article
Keywords:Brugia pahangi, L3, differentially expressed genes, troponin, tropomyosin, cuticular collagens
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Tetley, Dr Laurence and Devaney, Prof Eileen
Authors: Hunter, S.J., Martin, S.A., Thompson, F.J., Tetley, L., and Devaney, E.
Subjects:Q Science > QR Microbiology
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation > Bacteriology
College of Medical Veterinary and Life Sciences > School of Life Sciences > Life Sciences Infection Biology
Journal Name:Parasitology
Publisher:Cambridge University Press
ISSN:0031-1820
ISSN (Online):1469-8161
Published Online:8 September 2000
Copyright Holders:Copyright © 1999 Cambridge University Press
First Published:First published in Parasitology 119(2):189-198
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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