Production of biologically active human RelA (p65) in baculovirus-infected insect cells.
A recombinant baculovirus was constructed to express a cDNA encoding RelA (p65), a member of the NF-kappa B/Rel family of proteins. Infection of Spodoptera frugiderda insect cells with the recombinant baculovirus resulted in the production of the biologically active protein as measured by immunoblotting using RelA-specific antisera and by electrophoretic mobility shift assays. The recombinant protein bound specifically to an oligonucleotide containing the NF-kappa B consensus motif but not to that containing the unrelated Oct-1 consensus motif. Thus insect cell-derived RelA possess properties similar to the native protein and may be used in physical, biochemical, and pharmacological studies.
Chabot-Fletcher
M
M
Department of Immunopharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939, USA.
Ho
M L
ML
Breton
J J
JJ
Hanning
C R
CR
Amegadzie
B Y
BY
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology* / Q000378:metabolism; D051996:Transcription Factor RelA
D018076:DNA, Complementary; D016328:NF-kappa B; D011993:Recombinant Fusion Proteins; D051996:Transcription Factor RelA; D004247:DNA