Production of mouse granulocyte-macrophage colony-stimulating factor by gateway technology and transgenic rice cell culture

Yu Kuo Liu*, Li Fen Huang, Shin Lon Ho, Chun Yu Liao, Hsin Yi Liu, Ying Hui Lai, Su May Yu, Chung An Lu

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

28 Scopus citations

Abstract

To establish a production platform for recombinant proteins in rice suspension cells, we first constructed a Gateway-compatible binary T-DNA destination vector. It provided a reliable and effective method for the rapid directional cloning of target genes into plant cells through Agrobacterium-mediated transformation. We used the approach to produce mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) in a rice suspension cell system. The promoter for the αAmy3 amylase gene, which is induced strongly by sugar depletion, drove the expression of mGM-CSF. The resulting recombinant protein was fused with the αAmy3 signal peptide and was secreted into the culture medium. The production of rice-derived mGM-CSF (rmGM-CSF) was scaled up successfully in a 2-L bioreactor, in which the highest yield of rmGM-CSF was 24.6mg/L. Due to post-translational glycosylation, the molecular weight of rmGM-CSF was larger than that of recombinant mGM-CSF produced in Escherichia coli. The rmGM-CSF was bioactive and could stimulate the proliferation of a murine myeloblastic leukemia cell line, NSF-60.

Original languageEnglish
Pages (from-to)1239-1247
Number of pages9
JournalBiotechnology and Bioengineering
Volume109
Issue number5
DOIs
StatePublished - 05 2012

Keywords

  • Bioreactor
  • MGM-CSF
  • Recombinant protein production
  • Rice suspension cell
  • Sugar-depletion induced promoter

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