Haemonchus contortus: development of a two-step, differential-display PCR to detect differential gene expression in nematodes from immune and naïve sheep.
Haemonchus contortus is a blood-feeding nematode which parasitizes the abomasum of sheep and represents a serious constraint to sheep production. Anthelmintics are currently the most common method of worm control but the worldwide development of multiple-drug resistance and issues of residues in the food chain make alternatives to anthelmintics a priority. Biotechnology-driven solutions to parasitism include vaccines and silencing of genes regulating nematode development. To pursue gene targets that may be suitable for parasite control, a two stage differential-display PCR (dd-PCR) approach was developed to observe differential gene expression between Haemonchus from immune and control sheep. Twenty-four reproducible differentially-expressed bands were identified in 60 pairs of dd-PCR comparisons. The first of these cloned and sequenced corresponded to the H. contortus 60S ribosomal protein L35A. The remaining bands are being cloned and validated and may provide new targets for parasite control.
Rowe
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A
Faculty of Veterinary Science, McMaster Building (B14), University of Sydney, Camperdown Campus, NSW 2006, Australia. tonyr@vetsci.usyd.edu.au
Yun
K
K
Emery
D
D
Sangster
N
N
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000032:analysis; D005260:Female; D005786:Gene Expression Regulation; D006188:Haemonchiasis / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D006189:Haemonchus / Q000235:genetics* / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D016053:RNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D048348:Reverse Transcription; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000469:parasitology*; D016622:Silver Staining / Q000379:methods / Q000662:veterinary
D018076:DNA, Complementary; D016053:RNA, Protozoan