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Construction and evaluation of a chimeric protein made from Fasciola hepatica leucine aminopeptidase and cathepsin L1

Published online by Cambridge University Press:  02 October 2014

K. Hernández-Guzmán ,
A. Sahagún-Ruiz ,
A.J. Vallecillo ,
I. Cruz-Mendoza  and
H. Quiroz-Romero
K. Hernández-Guzmán
Affiliation:
Departamento de Parasitología
A. Sahagún-Ruiz
Affiliation:
Departamento de Microbiología e Inmunología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, C.P. 04510, MéxicoD.F., México
A.J. Vallecillo
Affiliation:
Escuela de Medicina Veterinaria y Zootecnia, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca, Ecuador
I. Cruz-Mendoza
Affiliation:
Departamento de Parasitología
H. Quiroz-Romero*
Affiliation:
Departamento de Parasitología
*
*Fax: +52-56225898 E-mail: [email protected], [email protected]
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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.

Type
Research Papers
Information
Journal of Helminthology , Volume 90 , Issue 1 , January 2016 , pp. 7 - 13
Copyright
Copyright © Cambridge University Press 2014 

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