title: Overexpression of soluble human thymosin alpha 1 in Escherichia coli.
abstract: Synthesized gene of human thymosin alpha 1 (Talpha1) was inserted into pET-28a, pET-9c, pThioHis B, pGEX-2T or pBV222 and then inductively expressed in strains of Escherichia coli. Among the five expression systems, the BL21/pET-28a system provides the highest expression level of fusion protein in a soluble form, which is up to 70% of total expressed bacterial proteins as visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The resulting fusion protein purified through nickel affinity chromatography accounts for 2.53% of the wet bacterial pellet weight and reaches 94.5% purity by SDS-PAGE. These results indicate the potential of this expression system for high-throughput production of recombinant Talpha1.
authors.affiliation: Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Pei-Fu
authors.initials: PF
authors.lastname: Chen
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Zhang
authors.forename: Geng-Feng
authors.initials: GF
authors.lastname: Fu
authors.forename: Gen-Xing
authors.initials: GX
authors.lastname: Xu
authors.forename: Ya-Yi
authors.initials: YY
authors.lastname: Hou
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics*; D012995:Solubility; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D013947:Thymosin; D000077596:Thymalfasin