AI Article Synopsis
- The study focuses on the gene Hc-daf-22 from the parasitic nematode Haemonchus contortus, which is significant due to its role in livestock health and its response to environmental stressors.
- Researchers used genetic techniques like genome walking and qRT-PCR to characterize Hc-daf-22, confirming its structure and function through experiments in C. elegans, a model organism.
- The findings suggest that Hc-daf-22 not only shares functional similarities with its counterpart Ce-daf-22 but may also play an important role in developmental processes, with implications for understanding parasite biology and improving livestock management.
Article Abstract
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.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4966567 | PMC |
| http://dx.doi.org/10.1186/s13071-016-1704-1 | DOI Listing |
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