We employed two complementary approaches to glo bal comparison of the H. contortus gene set, using the Inparanoid algorithm full article to look in detail at orthologs with C. elegans and P. pacificus, and OrthoMCL for a wider view of gene family evolution with other clade V nema todes. Of 5,937 orthology groups between C. elegans and H. contortus, 5,012 are one Inhibitors,Modulators,Libraries to one orthologs, while an additional 899 orthologs could be identified in H. contortus and P. pacificus but not C. elegans, suggest ing they have been lost in the C. elegans lineage. A number of orthology groups are significantly expanded in H. contortus, including a family of 180 Haemonchus paralogs to a single C. elegans gene that lacks any functional annotation.
Other expanded groups include genes with likely roles in parasitism, such as Inhibitors,Modulators,Libraries cysteine rich secreted proteins, together with a set of helicase domains that include some Inhibitors,Modulators,Libraries with predicted signal peptides. Global analy sis of the evolution of entire gene families across the clade V nematodes confirms this pattern of significant diversity within the clade, and allowed us to identify H. contortus genes lacking clear orthologs in other clade IV or V nematodes. This shows that the Haemonchus specific proteome is enriched in genes encoding polypeptides involved in proteolysis, neurotrans mission and carbohydrate metabolism, and in secreted proteins and those expressed in the cuticle. While some of these genes are explored in detail in the more focused ana lyses below, others may represent novel candidate genes involved in the host parasite interface.
Gene expression in parasite life stages and gut As H. contortus progresses through its lifecycle, it must adapt to Inhibitors,Modulators,Libraries different environments with differing food sources and energy requirements and this is reflected in differential gene expression. RNA seq was used to analyze gene expres sion in six parasite life stages and the adult female gut. Samples were Inhibitors,Modulators,Libraries made in triplicate from independent http://www.selleckchem.com/products/17-AAG(Geldanamycin).html batches of parasite material for every stage, allowing sta tistically robust comparison of relative gene expression between the stages. We found significant expression for 17,483 genes in total from the 6 life stages studied. with between 13,962 and 15,569 genes expressed in each stage. A total of 11,295 genes were significantly up or down regulated through the lifecycle and we used annotation with Gene Ontology terms to investigate their broad functions. Metabolic enzyme expression throughout the parasite lifecycle was ana lyzed in more detail and will be discussed separately.