Dataset Information

Identification and Characterization of a Differentially Expressed Gene (07E12) in the Infective Larvae of the Parasitic Nematode Ascaris suum.

ABSTRACT: BACKGROUND:Parasitic nematodes cause animal and human diseases of major socio-economic importance worldwide. The suppression of parasite development at particular developmental stages could provide an alternative approach for nematode control. In this study, Ascaris suum was used as a model system in the study of the differentially expressed genes in the infective L3 stage. METHODS:The gene (07E12) was screened and identified from the subtractive cDNA library for the infective larvae of Ascaris suum using real-time quantitative PCR. Then, the full-length cDNA of 07E12 was characterized by 3' and 5' rapid amplification of cDNA ends (RACE). The characteristics of the gene were further analyzed using bioinformatic analyses. RESULTS:The results showed that the gene 07E12 was differentially expressed in the third-stage larvae of A. suum and its expression level in the infective larvae was much higher than in other stages. It was shown that the gene 07E12 had 99% identity with the corresponding sequences of the A. suum whole genome shotgun sequence containing the homologous sequences with conserved sequences of Neuropeptide-Like Protein family member. Likewise, by performing BLASTN and BLASTP searches in the GenBank™, it was shown that this gene had 99 % identity with A. suum cre-nlp-2 protein. CONCLUSION:This gene 07E12 which is differentially expressed in the third-stage larvae of A. suum may encode a neuropeptide-like protein family member, a very important molecule in the process of infecting a host.

SUBMITTER: Huang C

PROVIDER: S-EPMC4386041 | biostudies-literature | 2014 Apr-Jun

REPOSITORIES: biostudies-literature

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Identification and Characterization of a Differentially Expressed Gene (07E12) in the Infective Larvae of the Parasitic Nematode Ascaris suum.

Huang Cuiqin C You Jingzhou J Nai Fangfang F

Iranian journal of parasitology 20140401 2

<h4>Background</h4>Parasitic nematodes cause animal and human diseases of major socio-economic importance worldwide. The suppression of parasite development at particular developmental stages could provide an alternative approach for nematode control. In this study, Ascaris suum was used as a model system in the study of the differentially expressed genes in the infective L3 stage.<h4>Methods</h4>The gene (07E12) was screened and identified from the subtractive cDNA library for the infective lar ...[more]

PMID: 25848387

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Project description:Differential transcription in Ascaris suum was investigated using a genomic-bioinformatic approach. A cDNA archive enriched for molecules in the infective third-stage larva (L3) of A. suum was constructed by suppressive-subtractive hybridization (SSH), and a subset of cDNAs from 3075 clones subjected to microarray analysis using cDNA probes derived from RNA from different developmental stages of A. suum. The cDNAs (n = 498) shown by microarray analysis to be enriched in the L3 were sequenced and subjected to bioinformatic analyses using a semi-automated pipeline (ESTExplorer). Using gene ontology (GO), 235 of these molecules were assigned to 'biological process' (n = 68), 'cellular component' (n = 50), or 'molecular function' (n = 117). Of the 91 clusters assembled, 56 molecules (61.5%) had homologues/orthologues in the free-living nematodes Caenorhabditis elegans and C. briggsae and/or other organisms, whereas 35 (38.5%) had no significant similarity to any sequences available in current gene databases. Transcripts encoding protein kinases, protein phosphatases (and their precursors), and enolases were abundantly represented in the L3 of A. suum, as were molecules involved in cellular processes, such as ubiquitination and proteasome function, gene transcription, protein-protein interactions, and function. In silico analyses inferred the C. elegans orthologues/homologues (n = 50) to be involved in apoptosis and insulin signaling (2%), ATP synthesis (2%), carbon metabolism (6%), fatty acid biosynthesis (2%), gap junction (2%), glucose metabolism (6%), or porphyrin metabolism (2%), although 34 (68%) of them could not be mapped to a specific metabolic pathway. Small numbers of these 50 molecules were predicted to be secreted (10%), anchored (2%), and/or transmembrane (12%) proteins. Functionally, 17 (34%) of them were predicted to be associated with (non-wild-type) RNAi phenotypes in C. elegans, the majority being embryonic lethality (Emb) (13 types; 58.8%), larval arrest (Lva) (23.5%) and larval lethality (Lvl) (47%). A genetic interaction network was predicted for these 17 C. elegans orthologues, revealing highly significant interactions for nine molecules associated with embryonic and larval development (66.9%), information storage and processing (5.1%), cellular processing and signaling (15.2%), metabolism (6.1%), and unknown function (6.7%). The potential roles of these molecules in development are discussed in relation to the known roles of their homologues/orthologues in C. elegans and some other nematodes. The results of the present study provide a basis for future functional genomic studies to elucidate molecular aspects governing larval developmental processes in A. suum and/or the transition to parasitism.

Project description:CircRNAs, a novel class of ncRNA family, are endogenous transcriptional products involved in various biological and physiological processes in plants and animals. However, almost no information is available for circRNAs of parasitic helminths. In the present study, the circRNAs repertoire was comprehensively explored in Haemonchus contortus, a blood-sucking parasitic nematode of ruminants. In total, 20073 circRNAs were identified and annotated from three key developmental stages/genders of H. contortus including the free-living infective third-stage larvae (L3, 18883), parasitic adult female (Af, 3491), and male worms (Am, 2550) via deep-sequencing technology and bioinformatic analysis. Among these identified circRNAs, 71% were derived from exonic regions of protein-coding genes. The number of circRNAs transcribed from the X chromosome (4704) was higher than that from Chromosome I-V (3143, 3273, 3041, 3030, 2882). The amount of highly expressed circRNAs in third-stage larvae was significantly more abundant than that in adult stage. 15948 and 16847 circRNAs were differentially expressed between Af and L3s and between Am and L3, respectively. Among them, 13409 circRNAs existed in both comparisons. Furthermore, 1119 circRNAs were differentially expressed between Af_and_Am. GO enrichment analysis indicated that source genes of circRNAs differentially expressed between Am and L3 as well as between Af and L3 were significantly enriched in many biological processes, primarily including signaling, signal transduction and cell communication terms. KEGG analysis revealed that parental genes of differentially expressed circRNAs were mainly related to metabolism (pyruvate metabolism, glycerophospholipid metabolism, and carbon metabolism), MAPK signaling pathway, and phosphatidylinositol signaling system. Moreover, many circRNAs contained one or more miRNA potential binding sites, suggesting that they could regulate gene expression at the post-transcriptional level. Furthermore, the correctness of head-to-tail back splicing site and alternative circularization events were verified by Sanger sequencing using both divergent and convergent primers. Finally, the reliability of RNA-Seq data and the resistance of circRNAs to RNase R digestion were confirmed by quantitative RT-PCR. Taken together, our findings provide a foundation for elucidating the regulatory mechanisms of circRNAs in H. contortus, which will advance the understanding of circRNAs in parasitic nematodes.

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Dataset Information

Identification and Characterization of a Differentially Expressed Gene (07E12) in the Infective Larvae of the Parasitic Nematode Ascaris suum.

Publications

Identification and Characterization of a Differentially Expressed Gene (07E12) in the Infective Larvae of the Parasitic Nematode Ascaris suum.

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