Expression of Recombinant Human Octamer-Binding Transcription Factor 4 in Rice Suspension Cells

Int J Mol Sci. 2021 Jan 30;22(3):1409. doi: 10.3390/ijms22031409.

Abstract

The rice cell suspension culture system is a good way to produce recombinant human proteins, owing to its high biosafety and low production cost. Human Octamer-binding Transcription Factor 4 (Oct4) is a fundamental transcription factor responsible for maintaining human pluripotent embryonic stem cells. Recombinant Oct4 protein has been used to induce pluripotent stem cells. In this study, recombinant Oct4 proteins are produced via a sugar starvation-inducible αAmy3/RAmy3D promoter-signal peptide-based rice recombinant protein expression system. Oct4 mRNAs accumulate in the transgenic rice suspension cells under sugar starvation. The Oct4 recombinant protein is detected in the transgenic rice suspension cells, and its highest yield is approximately 0.41% of total cellular soluble proteins after one day of sugar starvation. The rice cell-synthesized recombinant human Oct4 protein show DNA-binding activity in vitro, which implies that the protein structure is correct for enabling specific binding to the target DNA motif.

Keywords: human Oct4; recombinant human embryonic transcription factor; rice cell suspension culture system; αAmy3 promoter.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • Containment of Biohazards
  • Gene Expression Regulation, Plant
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / isolation & purification*
  • Octamer Transcription Factor-3 / metabolism
  • Oryza / cytology*
  • Oryza / genetics
  • Plant Proteins / genetics
  • Plants, Genetically Modified / cytology
  • Plants, Genetically Modified / genetics
  • Promoter Regions, Genetic / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • alpha-Amylases / genetics

Substances

  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Plant Proteins
  • Recombinant Proteins
  • alpha-Amylases