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 journalArticle

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 1 1994

Fingerprint

Recombinant proteins
ras Genes
Cytomegalovirus
Oncogenes
Recombinant Proteins
Interleukin-6
Plasmids
Tetrahydrofolate Dehydrogenase
Methotrexate
Actins
Animals
Complementary DNA
Genes
Productivity
Cells
Clone Cells
Experiments

All Science Journal Classification (ASJC) codes

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

Cite this

Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells. / Yano, Takahiro; Teruya, Kiichiro; Shirahata, Sanetaka; Watanabe, Junko; Osada, Kazuhiro; Tachibana, Hirofumi; Ohashi, Hideya; Kim, Eun Ho; Murakami, Hiroki.

In: Cytotechnology, Vol. 16, No. 3, 01.01.1994, p. 167-178.

Research output: Contribution to journalArticle

Yano, Takahiro ; Teruya, Kiichiro ; Shirahata, Sanetaka ; Watanabe, Junko ; Osada, Kazuhiro ; Tachibana, Hirofumi ; Ohashi, Hideya ; Kim, Eun Ho ; Murakami, Hiroki. / Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells. In: Cytotechnology. 1994 ; Vol. 16, No. 3. pp. 167-178.
@article{6c1075310b9b47f394e03fd8fa4e5123,
title = "Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells",
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.",
author = "Takahiro Yano and Kiichiro Teruya and Sanetaka Shirahata and Junko Watanabe and Kazuhiro Osada and Hirofumi Tachibana and Hideya Ohashi and Kim, {Eun Ho} and Hiroki Murakami",
year = "1994",
month = "1",
day = "1",
doi = "10.1007/BF00749904",
language = "English",
volume = "16",
pages = "167--178",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells

AU - Yano, Takahiro

AU - Teruya, Kiichiro

AU - Shirahata, Sanetaka

AU - Watanabe, Junko

AU - Osada, Kazuhiro

AU - Tachibana, Hirofumi

AU - Ohashi, Hideya

AU - Kim, Eun Ho

AU - Murakami, Hiroki

PY - 1994/1/1

Y1 - 1994/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0028693575&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028693575&partnerID=8YFLogxK

U2 - 10.1007/BF00749904

DO - 10.1007/BF00749904

M3 - Article

C2 - 7766145

AN - SCOPUS:0028693575

VL - 16

SP - 167

EP - 178

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 3

ER -