title: [Optimization of prokaryotic expression condition and purification of anti-cancer protein NOR1 in E.coli].
abstract: OBJECTIVE
  To optimize the induction condition of human NOR1 gene expression in E.coli. and purify NOR1 recombinant proteins.
  
  
  METHODS
  A full-length cDNA of human NOR1 was inserted into the corresponding region of pET28b expression vector to yield recombinant prokaryotic expression vector pET28b-NOR1. The prokaryotic expression vector pET28b-NOR1 was introduced into the bacterial host E.coli Rosettablue(DE3). Recombinant NOR1 protein was induced at different conditions. Induction condition was optimized to obtain high yield of recombinant protein. At last, the recombinant NOR1 protein was purified by Ni-IDE chromatography resin.
  
  
  RESULTS
  Recombinant NOR1 protein was induced by IPTG in a dose-dependent manner. Increase of kanamycin concentration and induction temperature resulted in high yield of recombinant protein. The most recombinant protein was found in inclusion bodies. The recombinant His-NOR1 protein was purified with Ni-IDE chromatography resin under denature condition.
  
  
  CONCLUSIONS
  IPTG, kanamycin concentration and temperature can affect the expression of recombinant NOR1 protein in pET28b system. High yield of recombinant NOR1 protein is achieved by inducing 1 mmol/L IPTG and 200 μg/mL kanamycin at 37 degree. Recombinant His-NOR1 protein with high purity is purified.
authors.affiliation: Cancer Research Institute, Central South University, Changsha 410078, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Xiang
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Li
authors.forename: Mei
authors.initials: M
authors.lastname: Yi
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Li
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Zeng
authors.forename: Guiyuan
authors.initials: G
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D026901:Membrane Transport Proteins / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D026901:Membrane Transport Proteins; C479458:OSCP1 protein, human; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D007544:Isopropyl Thiogalactoside