Expression of human beta-interferon in Namalwa KJM-1 which was adapted to serum-free medium

✍ Scribed by Hiromasa Miyaji; Tamio Mizukami; Shinji Hosoi; Seiji Sato; Nobuo Fujiyoshi; Seiga Itoh

• Publisher: Springer
• Year: 1990
• Tongue: English
• Weight: 566 KB
• Volume: 3
• Category: Article
• ISSN: 0920-9069
• DOI: 10.1007/bf00143675

✦ Synopsis

A Namalwa cell line, KJM-1, which was adapted to serum-free medium is thought to be a very useful host cell line for recombinant DNA technology. Thus, the utility of Namalwa KJM-1 for expression of foreign genes was examined. As a model system human beta-interferon (ILIFN) gene was engineered for expression in this cell line. For construction of the l3-IFN expression vector pSE1131-4, the expression vector pAGEl was constructed and used. It contains simian virus 40 (SV40) early promoter, the rabbit IJ-globin RNA processing signals for splicing and polyadenylation, and SV40 early RNA processing signal for polyadenylation.

In addition to the above transcription unit, pAGEl contains the ampicillinresistance gene and G418-resistance gene. They can confer ampicillin resistance to Escherichia coli (E. coli) and G418 resistance to animal cells. To introduce plasmid DNA into cells, electroporation is a useful method (Wong, 1982;Potter, 1984). We have established conditions for DNA-mediated transfection of Namalwa KJM-1 cell line by electroporation. Among pSElBl-4-introduced cells, clone l-3 was further examined for the expression of &IFN in serum-free medium. The production level of B-IFN was elevated with the increase of the cell density. The results indicated that the Namalwa KJM-1 cell line is useful for production of foreign gene products.