A tissue specific approach for analysis of membrane and secreted protein antigens from Haemonchus contortus gut and its application to diverse nematode species.
General methods to conduct tissue specific analysis are largely lacking for nematodes. An approach is described that focused on isolation of membrane and secreted protein genes from the gut of the parasitic nematode Haemonchus contortus. The approach capitalized on a monoclonal antibody that recognizes multiple membrane and secreted worm proteins. Polyclonal antisera made against these proteins were used to screen expression cDNA libraries made either from adult worm gut or whole worm. The genes identified encode predicted or known membrane and secreted proteins from gut, including a cysteine protease, a zinc metallopeptidase and a previously described GA1 protein. Another gene, Hc40, was isolated from the whole worm cDNA library and is nearly identical to a vaccine patent sequence pBTA879. Tissue analysis demonstrated the intended focus on membrane and secreted proteins from parasite gut was achieved. Proteins related to each of those described were identified from other nematode species through data base analysis. Additionally, this analysis led to (1) identification of homologues of each gene in C. elegans; (2) deduction of a dimorphic structure in the Hc40 protein; (3) recognition of both monomorphic and dimorphic families of Hc40-related proteins; and (4) detection of two apparent classes of transcripts (mep1a and mep1b) that would each encode a divergent version of the putative zinc metallopeptidase MEP1. The tissue specific approach and information base described should generally contribute to investigations on nutrient digestion and related secretory processes in nematode gut.
Rehman
A
A
Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA.
Jasmer
D P
DP
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis* / Q000235:genetics; D000953:Antigens, Protozoan; D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics; D027682:Cation Transport Proteins; D003546:Cysteine Endopeptidases / Q000032:analysis / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D005656:Fungal Proteins; D015723:Gene Library; D017238:Genes, Helminth / Q000235:genetics; D006189:Haemonchus / Q000276:immunology*; D015801:Helminth Proteins / Q000032:analysis / Q000235:genetics; D007422:Intestines / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D008666:Metalloendopeptidases / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D009348:Nematoda / Q000235:genetics; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000032:analysis / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D012756:Sheep; D013045:Species Specificity
D000947:Antigens, Helminth; D000953:Antigens, Protozoan; D002352:Carrier Proteins; D027682:Cation Transport Proteins; D018076:DNA, Complementary; D005656:Fungal Proteins; C100816:GA1 protein, Haemonchus contortus; C117854:Gcp7 protein, Haemonchus contortus; C117853:Hc40 protein, Haemonchus contortus; D015801:Helminth Proteins; C088951:MEP1 protein, S cerevisiae; D008565:Membrane Proteins; D015800:Protozoan Proteins; D029701:Saccharomyces cerevisiae Proteins; D003546:Cysteine Endopeptidases; C117855:MEP1 protein, Haemonchus contortus; D008666:Metalloendopeptidases