title: Functional over-expression of the Stm1 protein, a G-protein-coupled receptor, in Schizosaccharomyces pombe.
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.
authors.affiliation: Division of Genomics & Proteomics, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, Korea.
authors.forename: Kyung-Sook
authors.initials: KS
authors.lastname: Chung
authors.forename: Dong-Uk
authors.initials: DU
authors.lastname: Kim
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Ryoo
authors.forename: Eun-Jung
authors.initials: EJ
authors.lastname: Kang
authors.forename: Misun
authors.initials: M
authors.lastname: Won
authors.forename: Lila
authors.initials: L
authors.lastname: Kim
authors.forename: Young-Joo
authors.initials: YJ
authors.lastname: Jang
authors.forename: Pil-Jae
authors.initials: PJ
authors.lastname: Maeng
authors.forename: Sei-Chang
authors.initials: SC
authors.lastname: Kim
authors.forename: Hyang-Sook
authors.initials: HS
authors.lastname: Yoo
authors.forename: Kwang-Lae
authors.initials: KL
authors.lastname: Hoe
mesh_terms: 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
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; C439675:Stm1 protein, S pombe; D019204:GTP-Binding Proteins