title: [Construction and expression of a eukaryotic expression vector containing human CR2-Fc fusion protein].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector containing human complement receptor 2 (CR2)-Fc and express the CR2-Fc fusion protein in Chinese hamster ovary (CHO) cells.
  
  
  METHODS
  The extracellular domain of human CR2 and IgG1 Fc were respectively amplified, ligated and inserted into the eukaryotic expression vector PCI-neo. After verified by restriction enzyme digestion and sequencing, the recombinant plasmid was transfected into CHO K1 cells. The ones with stable expression of the fusion protein were obtained by means of G418 selection. The expression of the CR2-Fc fusion protein was detected and confirmed by SDS-PAGE and Western blotting.
  
  
  RESULTS
  Restriction enzyme digestion and sequencing demonstrated that the recombinant plasmid was valid. SDS-PAGE showed that relative molecular mass (Mr;) of the purified product was consistent with the expected value. Western blotting further proved the single band at the same position.
  
  
  CONCLUSIONS
  We constructed the eukaryotic expression vector of CR2-Fc/PCI-neo successfully. The obtained fusion protein was active and can be used for the further study of the role in HIV control.
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Li
authors.forename: Zhihao
authors.initials: Z
authors.lastname: Wu
authors.forename: Chuanfu
authors.initials: C
authors.lastname: Zhang
authors.forename: Leili
authors.initials: L
authors.lastname: Jia
authors.forename: Hongbin
authors.initials: H
authors.lastname: Song
authors.forename: Yuanyong
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006678:HIV / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D017464:Receptors, Complement 3d / Q000235:genetics* / Q000302:isolation & purification; D017452:Receptors, IgG / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification; D014162:Transfection
chemical_list: D017464:Receptors, Complement 3d; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins