[Construction of C3b(AN42) and OVA(257-264) fusion gene and functional analysis of the fusion protein].
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.
Xu
Gui-lian
GL
Institute of Immunology, Third Military Medical University, Chongqing 400038, China. immuno07@mail.immu.com.cn
Guo
Bo
B
Zheng
Ping
P
Yang
Fei
F
Zou
Qiang
Q
Wu
Yu-zhang
YZ
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
D003470:Culture Media; D004527:Egg Proteins; D016056:Immunodominant Epitopes; C110366:OVA-8; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D003179:Complement C3b; D010047:Ovalbumin