Immunological characterization of the sheep prion protein expressed as fusion proteins in Escherichia coli.
The prion protein (PrP) from sheep was produced in large quantities of entire protein in Escherichia coli after fusion with a carboxy-terminal hexahistidine sequence. In contrast, amino-terminal fusion with glutathione S-transferase (GST) revealed a high susceptibility toward cleavage of the protein. Both recombinant proteins were recognised, at variable levels, in Western blots using a panel of antibodies against the 40-56, 89-104, 98-113 and 112-115 sequences of the prion protein, similarly to the abnormal prion protein extracted from scrapie-infected sheep. Interestingly, monoclonal antibody 3F4 was found to react with these three proteins in Western blot.
Baron
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TG
Agence Française de Sécurité Sanitaire des Aliments, Lyon, France. t.baron@lyon.afssa.fr
Betemps
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Groschup
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Madec
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D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006639:Histidine; D011328:Prions / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012756:Sheep
D011328:Prions; D011993:Recombinant Fusion Proteins; D006639:Histidine; D005982:Glutathione Transferase