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

S J Hunter, Samuel Allen Moore Martin, F J Thompson, L Tetley, E Devaney

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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.

Original languageEnglish
Pages (from-to)189-198
Number of pages10
JournalParasitology
Volume119
Issue number2
DOIs
Publication statusPublished - Aug 1999

Keywords

  • Brugia pahangi
  • L3
  • differentially expressed genes
  • troponin
  • tropomyosin
  • cuticular collagens
  • 3RD stage larvae
  • trichostrongylus-colubriformis
  • caenorhabditis-elegans
  • gene-expression
  • collagen genes
  • Malayi
  • sequences
  • antigen
  • identification

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