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Biochemistry (Moscow)

, 74:75 | Cite as

High human GLUT1, GLUT2, and GLUT3 expression in Schizosaccharomyces pombe

  • Yuxin Yang
  • Zongli Hu
  • Zhizhao Liu
  • Yi Wang
  • Xuqing Chen
  • Guoping ChenEmail author
Article

Abstract

In this study, three subfamily members of the human 12-transmembrane-domain cell-surface receptors GLUT1, GLUT2, and GLUT3 were heterologously expressed in the fission yeast Schizosaccharomyces pombe utilizing GST-GLUT fusion proteins. These fusion proteins were driven by the full-length nmt1 promoter (Pnmt1) derived from S. pombe. The transcription levels of the GST-GLUT fusion proteins were very high upon induction by removing thiamine from the media. One-step purification of the recombinant fusion proteins was achieved by GST-affinity chromatography. Approximately 300 µg of highly purified fusion protein were obtained from 3 g of wet cell paste (1 liter of cell culture), indicating that human membrane proteins can be efficiently expressed and purified in the fission yeast. With its available extensive genetic information and ease of genetic manipulation, the fission yeast is potentially a highly efficient host to express eukaryotic membrane proteins.

Key words

GLUT Schizosaccharomyces pombe membrane protein expression GST-affinity chromatography 

Abbreviations

DAB

3,3′-diaminobenzidine

EMM

Edinburgh minimal media

GLUTs

facilitated glucose transporters

GST

glutathione S-transferase

PVDF

polyvinylidene difluoride

YES

yeast extract plus supplements medium

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • Yuxin Yang
    • 1
  • Zongli Hu
    • 1
  • Zhizhao Liu
    • 1
  • Yi Wang
    • 1
  • Xuqing Chen
    • 1
    • 2
  • Guoping Chen
    • 1
    Email author
  1. 1.College of BioengineeringChongqing UniversityShapingbaChina
  2. 2.Beijing Research Center of Agro-BiotechnologyBeijingChina

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