title: [Expression of TMP Fc in Pichia pastoris and identification of its biological activity].
abstract: The DNA coding for the fusion protein of thromobopoietin mimetic peptide (TMP) and human IgG1 Fc fragment was amplified from recombinant plasmid pET28a/TMPFc, inserted into pPICZalphaA and transformed into Pichia pastoris using electroporation. The recombinants of correct phenotype were identified after screening on MDH and MMH culture medium. The fusion gene was verified with PCR and western blot. MTT method was used to test the activity of TMPFc in promoting the growth of Ba/ F3-mpl cell. The TMPFc with a 64 000 molecular weight was a secretary protein in the system and its expression amounted to 65% of the total protein in the medium supernatant. The TMPFc showed a promotive effect on the growth of Ba/F3-mpl in vitro. A significant portion of the secretary protein existed as dimer, which provided material for studying the dimer in future.
authors.affiliation: Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China.
authors.forename: Xiang-Zhong
authors.initials: XZ
authors.lastname: Ye
authors.forename: Qiang
authors.initials: Q
authors.lastname: Guo
authors.forename: Chao
authors.initials: C
authors.lastname: Li
authors.forename: Feng-Yun
authors.initials: FY
authors.lastname: Liu
authors.forename: Du-Sheng
authors.initials: DS
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D007074:Immunoglobulin G / Q000235:genetics*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013926:Thrombopoietin / Q000235:genetics*
chemical_list: D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D013926:Thrombopoietin