Expression of ricin B chain in Spodoptera frugiperda.
DNA encoding ricin B chain was derived from preproricin genomic DNA and ligated into the baculovirus transfer vector, pAcGP67A. Co-transfection of Spodoptera frugiperda Sf9 cells with BaculoGold DNA was followed by limiting dilution purification of recombinant baculovirus. Infection of SF9 cells at a multiplicity of infection of 1 in the presence of 25 mM lactose produced 3 mg/l of soluble, glycosylated, 34 kDa protein immunoreactive with monoclonal and polyclonal antibodies to ricin B chain. The recombinant ricin B chain had similar lectin-binding activity to plant ricin B chain. The recombinant protein reassociated with ricin toxin A chain similarly to ricin toxin B chain and the recombinant heterodimers had similar cell cytotoxicity to ricin.
Department of Medicine, Medical University of South Carolina, Charleston.
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Frankel
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Roberts
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Gulick
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Afrin
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Vesely
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Willingham
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D037102:Lectins / Q000378:metabolism; D037121:Plant Lectins; D010944:Plants / Q000378:metabolism; D011994:Recombinant Proteins; D012276:Ricin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D018411:Spodoptera / Q000235:genetics*
D037102:Lectins; D037121:Plant Lectins; D011994:Recombinant Proteins; D012276:Ricin