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Structural and functional characterization of a novel gene,Hc-daf-22,from the strongylid nematode Haemonchus contortus
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نویسنده
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guo x. ,zhang h. ,zheng x. ,zhou q. ,yang y. ,chen x. ,du a.
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منبع
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parasites and vectors - 2016 - دوره : 9 - شماره : 1
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چکیده
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Background: the strongylid nematode haemonchus contortus is a parasite of major concern for modern livestock husbandry because hostile environmental conditions may induce diapause in the early fourth-stage larvae. methods: a new gene hc-daf-22 was identified which is the homologue of ce-daf-22 and human scpx. genome walking and race were performed to obtain the whole cdna and genomic sequence of this gene. using qrt-pcr with all developmental stages as templates to explore the transcription level and micro-injection was applied to confirm the promoter activity of the 5′-flanking region. overexpression,rescue and rna interference experiments were performed in n2,daf-22 mutant (ok 693) strains of c. elegans to study the gene function of hc-daf-22. results: the full length gene of hc-daf-22 (6,939 bp) contained 16 exons separated by 15 introns,and encoded a cdna of 1,602 bp (533 amino acids,estimated at about 59.3 kda) with a peak in l3 and l4 in transcriptional level. the hc-daf-22 protein was consisted of a 3-oxoacyl-coa thiolase domain and a scp2 domain and evolutionarily conserved. the 1,548 bp fragment upstream of the 5′-flanking region was confirmed to have promoter activity compared with 5′-flanking region of ce-daf-22. the rescue experiment by micro-injection of daf-22 (ok693) mutant strain showed significant increase in body size and brood size in the rescued worms with significantly reduced or completely absent fat granules confirmed by oil red o staining,indicating that hc-daf-22 could partially rescue the function of ce-daf-22. furthermore,rnai with hc-daf-22 could partially silence the endogenous ce-daf-22 in n2 worms and mimic the phenotype of daf-22 (ok693) mutants. conclusion: the gene hc-daf-22 was isolated and its function identified using c. elegans as a model organism. our results indicate that hc-daf-22 shared similar characteristics and function with ce-daf-22 and may play an important role in peroxisomal β-oxidation and the development in h. contortus. © 2016 the author(s).
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کلیدواژه
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Caenorhabditis elegans; Diapause; Haemonchus contortus; Hc-daf-22; β-oxidation
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آدرس
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college of animal sciences,zhejiang provincial key laboratory of preventive veterinary medicine,zhejiang university,hangzhou,310058, China, zhejiang center of animal disease control,hangzhou,310020, China, college of animal sciences,zhejiang provincial key laboratory of preventive veterinary medicine,zhejiang university,hangzhou,310058, China, faculty of life science and biotechnology,ningbo university,ningbo,315211, China, college of animal sciences,zhejiang provincial key laboratory of preventive veterinary medicine,zhejiang university,hangzhou,310058, China, college of animal sciences,zhejiang provincial key laboratory of preventive veterinary medicine,zhejiang university,hangzhou,310058, China, college of animal sciences,zhejiang provincial key laboratory of preventive veterinary medicine,zhejiang university,hangzhou,310058,china,institute of preventive veterinary medicine,college of animal sciences,zhejiang university,hangzhou,zhejiang,310058, China
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Authors
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