Abstract:

To construct the genetic engineering bacteria highly expressing fusion protein of glutathione-S-transferase ( GST ) and C-terminal 71 amino acids of fibroblast growth factor 23 ( FGF23CTR ),and to test the immunogenicity of the fusion protein and  to provide experimental data for FGF23 specific monoclonal antibodies and the development of diagnosis reagent for  chronic kidney disease.Methods After FGF23CTR gene fragment was obtained by PCR,it was fused with GST by expression vector pGEX-4T-1,then was transformed in E.coli BL21(DE3). By inducing with 1IPTG,the fusion proteins were expressed solubly.The fused protein was purified by Glutathione Sepharose4B.The purity of GST-FGF23CTR by SDS-PAGE was shown to be higher than 90%.The   Balb/C mice  were immuned   with fusion protein to pepare antiserum,then  ELISA was used to assay antiserum titer.Results The GST-FGF23CTR expression vector was constructed successfully.The purity of fusion protein was more than 90% after purification,and it positively reacted  to the commercialization of FGF23 polyclonal antibody.The ELISA result showed that the confirmed fusion protein had  good immunogenicity.Conclusion The genetic engineering bacteria of GST-FGF23CTR is successfully constructed and the purified protein has good immunogenicity.The fused protein can be used for the FGF23 monoclonal antibody preparation and research on the biological function of FGF23.

Key words: fibroblast growth factor 23;immunogenicity;genetic engineering bacteria