含绿色荧光蛋白及人胰岛素基因的真核表达载体的构建

目的构建含绿色荧光蛋白(GFP)基因与人胰岛素基因的重组真核表达载体。方法将人胰岛素基因插入到真核表达载体pIRES2-EGFP的EcoRⅠ和BamHⅠ位点中,构建重组质粒pIRES2-EGFP-INS。酶切鉴定,测序证实。结果重组真核表达载体pIRES2-EGFP-INS经限制性内切酶EcoRⅠ和BamHⅠ酶切,电泳后显示360bp的INS的目的片段和5.3kb的pIRES2-EGFP载体片段,进一步测序并证明重组质粒连接正确。结论成功构建了pIRES2-EGFP-INS重组质粒,为简便快速了解胰岛素基因的转染效率奠定了基础。

Construction of eukaryotic expression vector containing the human insulin and green fluoresent protein gene

Objective To construct eukaryotic expression vector containing the human insulin gene and green fluoresent protein gene. Methods The human insulin (INS) gene digested with EcoRⅠ and BamHⅠwas inserted into MCS sites of eukaryotic expression vector pIRES2-EGFP to construct pIRES2-EGFP-INS. Recombinant plasmid was verified by sequencing analysis. Results The recombinant eukaryotic expression vector pIRES2-EGFP-INS was digested with EcoRⅠand Bam HI, and electrophoresis of the digested products showed two fragments: 360 bp fragment and 5.3 kb fragment.Sequencing analysis showed that the recombinant plasmid was correct. Conclusion Construction of pIRES2-EGFP-INS is successful, which would benefit facilitating the human insulin gene′s delivery into cell.