title: Construction and evaluation of a chimeric protein made from Fasciola hepatica leucine aminopeptidase and cathepsin L1.
abstract: Leucine aminopeptidase (LAP) and cathepsin L1 (CL1) are important enzymes for the pathogenesis and physiology of Fasciola hepatica. These enzymes were analysed in silico to design a chimeric protein containing the most antigenic sequences of LAP (GenBank; AAV59016.1; amino acids 192-281) and CL1 (GenBank CAC12806.1; amino acids 173-309). The cloned 681-bp chimeric fragment (rFhLAP-CL1) contains 270 bp from LAP and 411 bp from CL1, comprising three epitopes, DGRVVHLKY (amino acids 54-62) from LAP, VTGYYTVHSGSEVELKNLV (amino acids 119-137) and YQSQTCLPF (amino acids 161-169) from CL1. The ~25 kDa rFhLAP-CL1 chimeric protein was expressed from the pET15b plasmid in the Rosetta (DE3) Escherichia coli strain. The chimeric protein rFhLAP-CL1, which showed antigenic and immunogenic properties, was recognized in Western blot assays using F. hepatica-positive bovine sera, and induced strong, specific antibody responses following immunization in rabbits. The newly generated chimeric protein may be used as a diagnostic tool for detection of antibodies against F. hepatica in bovine sera and as an immunogen to induce protection against bovine fasciolosis.
authors.affiliation: Departamento de Parasitología.
authors.forename: K
authors.initials: K
authors.lastname: Hernández-Guzmán
authors.affiliation: Departamento de Microbiología e Inmunología,Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México,C.P. 04510,MéxicoD.F.,México.
authors.forename: A
authors.initials: A
authors.lastname: Sahagún-Ruiz
authors.affiliation: Escuela de Medicina Veterinaria y Zootecnia, Facultad de Ciencias Agropecuarias,Universidad de Cuenca,Cuenca,Ecuador.
authors.forename: A J
authors.initials: AJ
authors.lastname: Vallecillo
authors.affiliation: Departamento de Parasitología.
authors.forename: I
authors.initials: I
authors.lastname: Cruz-Mendoza
authors.affiliation: Departamento de Parasitología.
authors.forename: H
authors.initials: H
authors.lastname: Quiroz-Romero
mesh_terms: D000818:Animals; D056668:Cathepsin L / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology; D000939:Epitopes / Q000032:analysis / Q000235:genetics / Q000276:immunology; D005210:Fasciola hepatica / Q000235:genetics* / Q000276:immunology; D005211:Fascioliasis / Q000469:parasitology / Q000662:veterinary*; D015870:Gene Expression; D015801:Helminth Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D007114:Immunization; D007931:Leucyl Aminopeptidase / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D008099:Liver / Q000201:enzymology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology
chemical_list: D000939:Epitopes; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D007931:Leucyl Aminopeptidase; D056668:Cathepsin L