Expression of ricin B chain in Spodoptera frugiperda

A. Frankel, H. Roberts, L. Afrin, J. Vesely, M. Willingham

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)787-794
Number of pages8
JournalBiochemical Journal
Volume303
Issue number3
DOIs
StatePublished - 1994

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