Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells

Takahiro Yano, Kiichiro Teruya, Sanetaka Shirahata, Junko Watanabe, Kazuhiro Osada, Hirofumi Tachibana, Hideya Ohashi, Eun Ho Kim, Hiroki Murakami

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

In order to enhance recombinant protein productivity in animal cells, we developed the oncogene activated production (OAP) system. The OAP system is based on the premise that oncogenes are able to enhance promoter activity. To this end, we constructed reported plasmids by fusing various promoters to the human interleukin-6 (hIL-6) cDNA, and the effector plasmids by inserting individual oncogenes, for example c-myc, c-fos, v-jun, v-myb and c-Ha-ras, downstream from the human cytomegalovirus immediate early (CMV) promoter. Results of transient expression experiments with BHK-21 cells suggest that the CMV promoter is the most potent promoter examined and that the ras product is able to transactivate the β-actin, CMV and SRα promoters. Recombinant BHK-21 cells producing hIL-6 under the control of the CMV promoter were contransfected with the ras oncogene and dihydrofolate reductase gene, then selected with 50 nM methotrexate to coamplify the ras oncogene. We were able to rapidly establish a stable and highly productive clone which exhibited a 35-times higher production rate as compared to the control value.

Original languageEnglish
Pages (from-to)167-178
Number of pages12
JournalCytotechnology
Volume16
Issue number3
DOIs
Publication statusPublished - Jan 1994

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

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