Objective: To construct and express a eukaryotic expression plasmid of mice FGFR1-MIP3?/Fc fusion gene,and detect its expression in the cells of 293 T which to provide laboratory data for the study of FGFR1-MIP3?/Fc fusion protein against tumor angiogenesis.Methods: The FGFR1-MIP3? fusion gene fragment was amplified by PCR with primers;After digesting by restriction endonuclease,the FGFR1-MIP3? fusion gene fragment was inserted into the eukaryotic expression vector of pc DNA3.1(+)/Fc(Mouse IgG2a)to construct the expression plasmid of FGFR1-MIP3?/Fc;After confirming by PCR,enzyme digestion analysis,and sequencing,the recombinant plasmid of FGFR1-MIP3?/Fc was transferred into 293 T cells and the expression of the fusion gene was identified by ELISA.Results: The digestion by PCR,restriction endonuclease and DNA sequencing demonstrated that the recombinant plasmid FGFR1-MIP3?/Fc fusion gene was correctly constructed;the plasmid could expressed FGFR1-MIP3?/Fc protein in 293 T cells by ELISA.Conclusion: The eukaryotic expression Plasmid of FGFR1-MIP3?/Fc was successfully constructed and expressed in 293 T cells. |