Optimal expression of GUS gene from methyl jasmonate-inducible promoter in high density culture of transformed tobacco cell line BY-2

Ken Ichiro Suehara, Shinji Takao, Kenzo Nakamura, Nobuyuki Uozumi, Takeshi Kobayashi

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

The optimal expression of the β-glucuronidase (GUS) gene was studied in a tobacco cell line, BY-2, that was transformed with the GUS gene under the control of the cathepsin D inhibitor (CDI) promoter. In batch culture, the optimal induction time was in the late growth phase, and the optimal concentration of methyl jasmonate (MJ) was 0.7 mM. In fed-batch culture, the GUS specific activity when MJ was added several times was about 1.7-fold that when MJ was added only once. However, a significant decrease in the growth rate was observed after MJ addition. During the fed-batch culture, no medium components were depleted, and the presence of inhibitory metabolite(s) was observed. To remove inhibitory metabolite(s) from the medium, filtration culture was carried out, which gave a cell growth rate faster than that of the fed-batch culture. The final cell concentration and total GUS activity reached 480 g-fresh weight/l and 5.2 μkat/l, which were 1.3-fold and 1.5- fold the amounts obtained in fed-batch culture. The GUS specific activity using the CDI promoter was about 17-fold that obtained in a rbcS-promoter system studied previously. Efficient foreign gene production from transformed BY-2 cells was thus performed on a fermentor scale.

Original languageEnglish
Pages (from-to)51-55
Number of pages5
JournalJournal of Fermentation and Bioengineering
Volume82
Issue number1
DOIs
Publication statusPublished - 1996 Jan 1
Externally publishedYes

Keywords

  • CDI promoter
  • fed-batch culture
  • filtration culture
  • high cell density
  • tobacco cell
  • transgenic plant

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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