LMP3-EGFP融合蛋白真核表达载体的构建及其在HEK293细胞中的表达

为构建人LMP3-EGFP融合蛋白真核表达载体，并检测其在HEK293细胞中的表达及定位，通过基因重组的方法构建LMP3／pEGFP-N3重组真核表达载体，并通过酶切和基因测序鉴定。脂质体法转染HEK293细胞，用倒置荧光显微镜检测、分析其在HEK293细胞表达及定位。经酶切和基因测序鉴定LMP3／pEGFP-N3重组真核表达载体构建成功。转染HEK293细胞后，荧光显微镜检测显示融合蛋白仅在细胞浆中表达。说明基因重组技术可成功构建LMP3／pEGFP-N3重组体真核表达载体，重组表达的融合蛋白仅存在于细胞浆内。

Construction Eukaryotic Expression Vector of LIM Mineralization Protein 3 and Its Expression in HEK293 Cells

To construct the recombined eukaryotic expression vector of LMP3 /pEGFP-N3 and investigate the expression of fusion protein in HEK293 cells, the recombinant vector constructed by gene recombinant technology was analyzed by restriction enzyme digestion and DNA sequencing. The recombinant vector was transfected into HEK293 cells with liposome transfection reagents for transient expression. The expression of chimeric protein LMP3-EGFP was identified by fluorescent microscope. The recombinant plasmid proved successful by restriction enzyme digestion and DNA sequencing. The expression of the chimeric protein was shown by fluorescent microscope. The chimeric protein only existed in cytoplasm. It is conclused that the recombinant eukaryotic vector of LMP3/pEGFP-N3 is successfully constructed by the gene recombinant technology and the fusion protein only exists in cytoplasm.