title: Expression, purification, and characterization of recombinant human keratinocyte growth factor-2 in Pichia pastoris.
abstract: Keratinocyte growth factor-2 (KGF-2) is a member of the fibroblast growth factor family. The full-length human KGF-2 coding sequence, gained by synthesizing, was cloned into the pPICZalphaA vector in frame with the yeast alpha-factor secretion signal under the transcriptional control of the AOX promoter and integrated into Pichia pastoris strain GS115. In shake-flask culture induced with methanol, the rhKGF-2 content was about 17.5% of the total secreted proteins. Under the optimal conditions, stable production of rhKGF-2 around 1.0g/l was achieved. The recombinant protein was purified by heparin affinity chromatography. A preliminary biochemical characterization of purified rhKGF-2 was performed both by Western blot analysis and biological activity analysis, and the result demonstrated that the recombinant KGF-2 was expressed successfully.
authors: Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors: Yanchun
authors: Y
authors: Wang
authors: Shengling
authors: S
authors: Yuan
authors: Peng
authors: P
authors: Wang
authors: Xiangxin
authors: X
authors: Liu
authors: Dewen
authors: D
authors: Zhan
authors: Zhaoshan
authors: Z
authors: Zhang
mesh_terms: D001483:Base Sequence; D001709:Biotechnology; D015153:Blotting, Western; D017931:DNA Primers / Q000235:genetics; D051526:Fibroblast Growth Factor 10 / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D051526:Fibroblast Growth Factor 10; D011994:Recombinant Proteins