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  • Immunoprecipitation of Fasciola hepatica mRNA in vitro translation products using infection and hyperimmune sera.

Immunoprecipitation of Fasciola hepatica mRNA in vitro translation products using infection and hyperimmune sera.

The American journal of tropical medicine and hygiene (1988-05-01)
G V Hillyer, D W Taylor
ABSTRACT

Total RNA containing messenger RNA has been isolated from adult Fasciola hepatica and translated in vitro using the rabbit reticulocyte lysate system. L-[35S]-methionine labeled translation products have been immunoprecipitated with sera collected from rabbits infected with F. hepatica, rabbits immunized with adult worm homogenate, and a rabbit and 5 calves immunized with a Fasciola/Schistosoma cross-reactive, cross-protective antigen, denoted FhSmIII(M). Precipitated antigens were analyzed by SDS-polyacrylamide gel electrophoresis. The major antigens recognized by hyperimmune sera had apparent molecular weights of 12,500, 13,500, 14,000, 26,500, and 27,000. The rabbit anti-FhSmIII(M) serum precipitated a 13,500 Mr antigen. Sera collected from calves immunized with FhSmIII(M) also precipitated a 13,500 Mr antigen together with a 26,500 Mr molecule. Antibody with specificity for F. hepatica mRNA translation products could be detected in rabbit serum 2 weeks after infection.

MATERIALS
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Millipore
Protein A–Sepharose 6MB, aqueous ethanol suspension