Abstract

The human CCR5, a subfamily of G-protein-coupled, seven-transmembrane-domain cell-surface chemokine receptors that binds to β-chemokine, was heterologously expressed in fission yeast, Schizosaccharomyces pombe, through CCR5-GST fusion protein. The CCR5-GST fusion protein was driven by the full-length nmt1 promoter (Pnmt1) derived from S. pombe. The transcription level of CCR5-GST fusion protein was very high when induced by deprivation of thiamine in the media. About 200 μg of highly purified CCR5-GST fusion protein was obtained from 3 g of wet cell paste (one liter of cell culture). The SDS-PAGE and western blot analysis indicated that the human membrane protein was efficiently expressed and purified infission yeast.

Key words: Human chemokine receptor CCR-5, fission yeast, membrane protein expression, affinity chromatography.