Production of functional S-II in Bombyx mori cells.
A cDNA for murine transcription factor S-II (Hirashima et al., J. Biol. Chem (1988) 263, 3858-3863) was inserted into a silkworm baculovirus vector and expressed in Bm-N cells, a cell line of Bombyx mori maintained in the laboratory. Recombinant S-II was purified from virus-infected cell extracts to near homogeneity by ammonium sulfate fractionation, and chromatographies on DEAE-cellulose and phosphocellulose. About 60 micrograms of recombinant S-II was obtained from 1 g of virus-infected cells. The molecular mass and specific activity of recombinant S-II were exactly the same as those of authentic S-II purified from Ehrlich ascites tumor cells.
Hirashima
S
S
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Horikoshi
N
N
Sekimizu
K
K
Natori
S
S
D000818:Animals; D015153:Blotting, Western; D012831:Bombyx / Q000378:metabolism*; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology; D002460:Cell Line; D002845:Chromatography; D003001:Cloning, Molecular; D004257:DNA Polymerase II / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005590:Fractional Precipitation; D005822:Genetic Vectors; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; D014162:Transfection; D014780:Viruses / Q000235:genetics
D009363:Neoplasm Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; D004257:DNA Polymerase II