【摘要】 目的构建小鼠成纤维细胞生长因子受体1/巨噬细胞炎性蛋白3a-Fc融合基因(FGFR1/MIP3a-Fc)的真核表达质粒,并在293T细胞中的表达。方法设计合成FGFR1/MIP3a融合基因引物,用PCR方法扩增获得FGFR1/MIP3a基因片段,用内切酶消化后插入pc DNA3.1(+)/Fc(Mouse Ig G2a)质粒中,构建FGFR1/MIP3a-Fc融合基因表达质粒;经PCR鉴定、酶切鉴定以及测序鉴定后,将该融合基因表达质粒瞬时转染293T细胞,用ELISA法检测其在293T细胞的表达。结果经PCR、酶切鉴定以及测序证实,FGFR1/MIP3a-Fc融合基因表达质粒构建成功;ELISA检测FGFR1/MIP3a-Fc融合基因表达质粒能够在293T细胞中表达。结论 FGFR1/MIP3a-Fc融合基因表达质粒构建成功,该质粒能够在293T细胞中表达。更多还原

【Abstract】 Objective To construct and express a eukaryotic expression plasmid of mice FGFR1/MIP3a-Fc fusion gene,and detect its expression in the cells of 293 T. Methods The FGFR1/MIP3 a fusion gene fragment was amplified by PCR with primers; After digesting by restriction endonuclease, the FGFR1/MIP3 a fusion gene fragment was inserted into the eukaryotic expression vector of pc DNA3.1(+)/Fc(Mouse Ig G2a) to construct the expression plasmid of FGFR1/MIP3a-Fc; After confirming by PCR, enzyme digestion analysis, and sequencing, the recombinant plasmid of FGFR1/MIP3a-Fc was transferred into 293 T cells and the expression of the fusion gene was identified by ELISA. Results The identification of PCR and restriction endonuclease and DNA sequencing demonstrated that the recombinant plasmid FGFR1/MIP3a-Fc fusion gene was correctly constructed; the expression of FGFR1/MIP3a-Fc protein in 293 T cells was confirmed by ELISA. Conclusion The eukaryotic expression plasmid of FGFR1/MIP3a-Fc was successfully constructed, which can be expressed in 293 T cells.更多还原