Abstract
We report here the first functional over-expression of the Stm1 protein, a G-protein-coupled receptor with seven-trans-membrane spanning regions, in a homologous expression system without internal modification of the open reading frame of Stm1. The entire coding sequence, except for the termination codon followed by a C-terminal His6 tag, has been cloned into the pREP1 vector. The functionally active Stm1-His6 was over-expressed in Schizosaccharomyces pombe under the control of the nmt1 (no message in thiamine) promoter. The expression after induction was 120 times as much as that of control before induction and it gave approximately 500 ng protein/2 x 10(7) cells.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Escherichia coli / genetics
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Escherichia coli / metabolism
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GTP-Binding Proteins / biosynthesis
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GTP-Binding Proteins / genetics
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Gene Expression Regulation, Fungal / genetics*
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Membrane Proteins / biosynthesis*
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Membrane Proteins / genetics*
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Schizosaccharomyces / genetics*
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Schizosaccharomyces / metabolism*
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Schizosaccharomyces pombe Proteins / biosynthesis*
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Schizosaccharomyces pombe Proteins / genetics*
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Transformation, Genetic
Substances
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Membrane Proteins
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Recombinant Proteins
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Schizosaccharomyces pombe Proteins
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Stm1 protein, S pombe
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GTP-Binding Proteins