title: [Construction of C3b(AN42) and OVA(257-264) fusion gene and functional analysis of the fusion protein].
abstract: OBJECTIVE
  To explore the role of complement system in the T cell activation.
  
  
  METHODS
  OVA(257-264) CTL epitope and C3b(AN42) encoding genes were synthesized and cloned into expression vector pEGFP-N1. COS-7 cells were transfected with the recombinant plasmid pEGFP-N1. The expression of the fusion protein was determined by FACS. Twenty-four hours after transfection, cultural supernatants of the transfected cells were collected and added to Ana-1 cell culture. Laser confocal microscopy was used to detect the binding and presentation of the fusion protein by Ana-1 cells.
  
  
  RESULTS
  C3b(AN42)-OVA(257-264) fusion protein was expressed in COS-7 cells. The fusion protein could bind to Ana-1 cells and OVA(257-264) CTL epitope was cross-presented on the surface of Ana-1 cells.
  
  
  CONCLUSIONS
  The C3b(AN42)-OVA(257-264) fusion protein can express in COS-7 cells and possesses natural biological activity.
authors.affiliation: Institute of Immunology, Third Military Medical University, Chongqing 400038, China. immuno07@mail.immu.com.cn
authors.forename: Gui-lian
authors.initials: GL
authors.lastname: Xu
authors.forename: Bo
authors.initials: B
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Zheng
authors.forename: Fei
authors.initials: F
authors.lastname: Yang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zou
authors.forename: Yu-zhang
authors.initials: YZ
authors.lastname: Wu
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D003179:Complement C3b / Q000235:genetics* / Q000378:metabolism; D045142:Cross-Priming; D003470:Culture Media; D004527:Egg Proteins / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D016056:Immunodominant Epitopes / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D010047:Ovalbumin / Q000235:genetics* / Q000378:metabolism; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D003470:Culture Media; D004527:Egg Proteins; D016056:Immunodominant Epitopes; C110366:OVA-8; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D003179:Complement C3b; D010047:Ovalbumin