Functional over-expression of the Stm1 protein, a G-protein-coupled receptor, in Schizosaccharomyces pombe.
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.
Chung
Kyung-Sook
KS
Division of Genomics & Proteomics, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, Korea.
Kim
Dong-Uk
DU
Ryoo
Sung-Woo
SW
Kang
Eun-Jung
EJ
Won
Misun
M
Kim
Lila
L
Jang
Young-Joo
YJ
Maeng
Pil-Jae
PJ
Kim
Sei-Chang
SC
Yoo
Hyang-Sook
HS
Hoe
Kwang-Lae
KL
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014170:Transformation, Genetic
D008565:Membrane Proteins; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; C439675:Stm1 protein, S pombe; D019204:GTP-Binding Proteins