title: Expression, purification and characterization of a recombinant Tat47-57-Oct4 fusion protein in Pichia pastoris.
abstract: The transcription factor, Oct-4, is involved in the self-renewal of undifferentiated embryonic stem cells, and is also significant in the reprogramming process and in the development of tumors. In the present study, the fusion protein, Tat47‑57-Oct4, was secreted by the signal peptide of human serum albumin in Pichia pastoris under the control of alcohol oxidase promoter 1. The yield of recombinant Tat47‑57-Oct4 fusion protein was ~210 mg/l. Following pilot‑scale fermentation, Tat47‑57-Oct4 was purified by ammonium sulfate precipitation, Vivaflow 200 ultrafiltration and SP Sepharose fast flow chromatography in order to obtain 95.6% purity. Immunofluorescence analysis validated the ability of Tat47‑57-Oct4 to cross the cell membrane. The results demonstrated that the experimental procedure developed in the present study could produce large quantities of active Tat47‑57-Oct4 fusion protein from P. pastoris.
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Haotian
authors: H
authors: Wang
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Xinmin
authors: X
authors: Zhang
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Ning
authors: N
authors: Kong
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Anhui
authors: A
authors: Wei
authors: College of Basic Medical Science of China Medical University, Shenyang, Liaoning 110001, P.R. China.
authors: Yanhong
authors: Y
authors: Zhang
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Jie
authors: J
authors: Ma
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Yulai
authors: Y
authors: Zhou
authors: School of Pharmaceutical Sciences of Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Weiqun
authors: W
authors: Yan
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D003001:Cloning, Molecular; D005285:Fermentation; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012709:Serum Albumin / Q000235:genetics*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
chemical_list: D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D054322:tat Gene Products, Human Immunodeficiency Virus