title: Characterisation of the two most abundant genes in the Haemonchus contortus expressed sequence tag dataset.
abstract: Analysis of the Haemonchus contortus Expressed sequence tag (EST) dataset revealed that almost 10% of all ESTs (1719 ESTs) belong to a family of related genes. Close analysis of the ESTs suggests that these represent two genes (called here Hc-nim-1 and Hc-nim-2) with multiple alleles of each. These genes show significant similarity to two genes from Caenorhabditis elegans, F54D5.3 (Wormbase accession WBGene00010049, corresponding protein WP:CE28033) and F54D5.4 (WBGene00010050, WP:CE03409) of unknown function. Reverse transcriptase coupled-PCR showed that both genes are transcribed from the L4 stage onwards and are transcribed in both male and female adult worms. A partial bacterial recombinant of the Hc-NIM-1 protein was made and used to raise antiserum in rabbits which recognised a 19 kDa antigen in the water soluble protein fraction of adult worms. By immunohistochemistry, the Hc-NIM-1 protein was localised in the hypodermis of the pharyngeal region of adult worms but not posterior in the hypodermis surrounding the reproductive tract. To investigate the function of this novel protein family we conducted a RNA interference experiment for the homologuous proteins in C. elegans. No visible phenotype was detected after simultaneous RNAi treatment for both Ce-F54D5.3 and Ce-F54D5.4.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, UK.
authors.forename: P
authors.initials: P
authors.lastname: Geldhof
authors.forename: C
authors.initials: C
authors.lastname: Whitton
authors.forename: W F
authors.initials: WF
authors.lastname: Gregory
authors.forename: M
authors.initials: M
authors.lastname: Blaxter
authors.forename: D P
authors.initials: DP
authors.lastname: Knox
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth; D001483:Base Sequence; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D020224:Expressed Sequence Tags; D005260:Female; D017238:Genes, Helminth; D006189:Haemonchus / Q000235:genetics*; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D011817:Rabbits; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
chemical_list: D000947:Antigens, Helminth; D029742:Caenorhabditis elegans Proteins; D015801:Helminth Proteins