title: Expression, purification, and secondary structure characterization of recombinant KCTD1.
abstract: Potassium channel tetramerization domain containing 1 (KCTD1) contains a BTB domain, which can facilitate protein-protein interactions that may be involved in the regulation of signaling pathways. Here we describe an expression and purification system that can provide a significant amount of recombinant KCTD1 from Escherichia coli. The cDNA encoding human KCTD1 was amplified and cloned into the expression vector pET-30a(+). The recombinant protein was expressed in E. coli BL21(DE3) cells and subsequently purified using affinity chromatography. To confirm that KCTD1 was correctly expressed and folded, the molecular weight and conformation were analyzed using mass spectroscopy, Western blot, and circular dichroism. Optimizing KCTD1 expression and investigating its secondary structure will provide valuable information for future structural and functional studies of KCTD1 and KCTD family proteins.
authors.affiliation: The State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Fanghua
authors.initials: F
authors.lastname: Mei
authors.forename: Jin
authors.initials: J
authors.lastname: Xiang
authors.forename: Song
authors.initials: S
authors.lastname: Han
authors.forename: Yuan
authors.initials: Y
authors.lastname: He
authors.forename: Yajing
authors.initials: Y
authors.lastname: Lu
authors.forename: Jian
authors.initials: J
authors.lastname: Xu
authors.forename: Deyin
authors.initials: D
authors.lastname: Guo
authors.forename: Gengfu
authors.initials: G
authors.lastname: Xiao
authors.forename: Po
authors.initials: P
authors.lastname: Tien
authors.forename: Guihong
authors.initials: G
authors.lastname: Sun
mesh_terms: D002942:Circular Dichroism; D056970:Co-Repressor Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D006801:Humans; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
chemical_list: D056970:Co-Repressor Proteins; C529677:KCTD1 protein, human; D011994:Recombinant Proteins; D012097:Repressor Proteins