title: [Cloning and expressing of golgi protein73 gene fragment and preparation of monoclonal antibodies against the recombinant protein].
abstract: OBJECTIVE
  To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein.
  
  
  METHODS
  GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot was used to detect specificity of mAbs.
  
  
  RESULTS
  The prokaryotic plasmid expressing the recombinant protein was constructed, and the GP73 recombinant protein was expressed and purified. Five hybridoma cell lines that secreted anti-GP73 mAbs were obtained. 2 of 5 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with GP73 protein.
  
  
  CONCLUSIONS
  The GP73 recombinant protein is highly purified and has strong antigenicity. The anti-GP73 mAbs were prepared successfully.
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Jing-Xia
authors.initials: JX
authors.lastname: Guo
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Hong-Shan
authors.initials: HS
authors.lastname: Wei
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Yong-Ji
authors.initials: YJ
authors.lastname: Song
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Ai-Xia
authors.initials: AX
authors.lastname: Liu
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Bo-An
authors.initials: BA
authors.lastname: Li
authors.affiliation: Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
authors.forename: Yuan-Li
authors.initials: YL
authors.lastname: Mao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D003001:Cloning, Molecular; D015870:Gene Expression; D056945:Hep G2 Cells; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; C410824:GOLM1 protein, human; D008565:Membrane Proteins; D011994:Recombinant Proteins