title: [Expression of glutathione S-transferase-aquaporin 1 carboxyl terminal domain fusion protein in Escherchia coli].
abstract: We constructed a recombinant plasmid of water channel protein Aquaporin 1 (AQP1) carboxyl terminal domain (DNA sequence from 700bp-801bp) in pGEX-4T-1 vector and express the carboxyl terminal hydrophilic peptide AQP1 in E. coli. In this study, the DNA sequence of AQP1 hydrophilic peptide was amplified by PCR and was cloned into pGEX-4T-1 expression vector. After identified by restriction enzyme digestion and sequencing, the recombinant clone was transformed into the competent expression cells of E. coli BL21. The GST-AQP1 fusion protein was induced by IPTG and further purified by Glutathione Sepharose 4B to obtain a fusion protein with molecular weight of 30KD. So the fusion protein of AQP1 C-terminal hydrophilic peptide combined with GST was successfully expressed and purified. We set up important bases for the further research in AQP1 gene function.
authors.affiliation: Membrane Channel Research Laboratory, School of Life Sciences, Northeast Normal University, Changchun 130024. lixm441@nenu.edu.cn
authors.forename: Xiao Meng
authors.initials: XM
authors.lastname: Li
authors.forename: Jiang
authors.initials: J
authors.lastname: Li
authors.forename: Nan Yang
authors.initials: NY
authors.lastname: Yang
authors.forename: Xin Gang
authors.initials: XG
authors.lastname: Guan
authors.forename: Shu Zhi
authors.initials: SZ
authors.lastname: Zhang
authors.forename: Wei Li
authors.initials: WL
authors.lastname: Hui
authors.forename: Tong Hui
authors.initials: TH
authors.lastname: Ma
mesh_terms: D051398:Aquaporin 1 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D051398:Aquaporin 1; D005982:Glutathione Transferase