Expression of ricin B chain in Spodoptera frugiperda.
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ABSTRACT: 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.
SUBMITTER: Frankel A
PROVIDER: S-EPMC1137616 | biostudies-other | 1994 Nov
REPOSITORIES: biostudies-other
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