Characterisation and expression of an Hsp70 gene from Parastrongyloides trichosuri.
Parastrongyloides trichosuri is a nematode parasite of Australian brushtail possums that has an alternative free-living life cycle which can be readily maintained indefinitely in a laboratory setting. The ability to maintain this parasite in a free-living cycle and induce it to parasitism at the free-living L1 stage makes this an excellent model for the study of genes associated with parasitism. A 70kD protein from infective larvae of P. trichosuri that appears to be immunogenic in infected possums has been identified as a heat shock protein (Hsp)70 homologue. The complete gene for Pt-Hsp70 was cloned and sequenced. The protein encoded by the Pt-Hsp70 gene is the likely orthologue of the Caenorhabditis elegans protein, Hsp70A, also known as hsp-1. Reverse transcriptase-PCR data indicate that Pt-Hsp70 (designated Pt-hsp-1) is expressed at readily detectable levels in all developmental stages of both the parasitic and free-living P. trichosuri life cycles and the promoter is mildly inducible by heat shock. Bioinformatic analysis of expressed sequence tag databases indicates that C. eleganshsp-1 homologues, together with C. eleganshsp-3 homologues, are the predominant members of the Hsp70 superfamily that are normally expressed in parasitic stages of the Strongyloididae family. Promoter fusions to a beta-galactosidase coding sequence were prepared and introduced into wild type C. elegans to produce transgenic nematodes. Reporter gene expression was clearly present within embryonic cells and within intestinal cells of larval and adult stages. Thus, the expression of the Pt-hsp-1 promoter within P. trichosuri and transgenic C. elegans appears similar to the known expression of C. elegans hsp-1. This promoter should be of value in efforts to develop genetic manipulation tools for P. trichosuri.
Newton-Howes
J
J
AgResearch Ltd, Wallaceville Animal Research Centre, Ward Street, P.O. Box 40063, Upper Hutt, New Zealand.
Heath
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Shoemaker
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CB
Grant
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WN
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D019295:Computational Biology; D018507:Gene Expression Regulation, Developmental; D017238:Genes, Helminth; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000302:isolation & purification; D015801:Helminth Proteins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013321:Strongyloides / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D013322:Strongyloidiasis / Q000469:parasitology; D014170:Transformation, Genetic; D048330:Trichosurus / Q000469:parasitology
D029742:Caenorhabditis elegans Proteins; D018840:HSP70 Heat-Shock Proteins; D015801:Helminth Proteins