title: Protein expression of human neuron-specific enolase and its antiserum preparation.
abstract: OBJECTIVE
  To clone and express human neuron-specific enolase (HuNSE) protein and prepare NSE-specific antibody for prion disease diagnosis.
  
  
  METHODS
  HuNSE gene was amplified by RT-PCR and subcloned into a HIS-tagged expression vector pQE30 after sequence verification. HIS-NSE fusion protein expression was obtained in E. coli M15 after IPTG induction followed by purification of the fusion protein by Ni-NTA affinity chromatography. Two male rabbits were immunized for 4 times with the purified protein, and the antiserum against NSE protein was collected and evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohistochemistry.
  
  
  RESULTS
  SDS-PAGE assay yielded an approximately 22 kD HIS-NSE fusion protein. The prepared antiserum could recognize both recombinant NSE protein and native NSE protein extracted from the brain tissues of different mammalian species as shown by Western blotting and immunohistochemistry.
  
  
  CONCLUSIONS
  High expression of HuNSE is obtained in E. coli and the prepared antiserum against HuNSE can be used potentially for diagnosis of prion-associated diseases and other nervous degeneration diseases.
authors.affiliation: Institue of Neurobiology, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. xxli-2001@126.com
authors.forename: Xin-li
authors.initials: XL
authors.lastname: Xiao
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Xin-lin
authors.initials: XL
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Hui-ying
authors.initials: HY
authors.lastname: Jiang
authors.forename: Bao-yun
authors.initials: BY
authors.lastname: Zhang
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000097:blood / Q000276:immunology*; D007150:Immunohistochemistry; D008297:Male; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D017096:Prion Diseases / Q000175:diagnosis* / Q000276:immunology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D010751:Phosphopyruvate Hydratase