人Elf5基因真核表达质粒的构建及其亚细胞定位

摘要

Objective:To construct the eukaryotic pEGFP-hElf5 and identify the subcellular localization of Elf5 in MCF7 cells. Methods:Elf5 fragment was subcloned into pEGFP-C1 after PCR. The plasmid was verified by en-zyme digestion,DNA sequencing and its expression was subsequently checked by Western blot. Immunofluorescence was used to identify its subcellular location. Results:Elf5 was subcloned into pEGFP-C1 successfully. Conclusion:Eukaryotic pEGFP-hElf5 was constructed successfully and Elf5 locates in cell nucleus.%目的：构建人Elf5真核细胞表达质粒，并研究其在乳腺癌MCF7细胞中的亚细胞定位。方法：PCR扩增人Elf5基因，将其克隆入真核表达载体pEGFP-C1中，构建含人Elf5基因的真核表达质粒pEGFP-hElf5。利用酶切鉴定，DNA测序，免疫印迹及免疫荧光方法鉴定Elf5真核表达质粒及Elf5蛋白的亚细胞定位。结果：经酶切鉴定、DNA测序、免疫印迹方法确认Elf5基因成功导入真核表达载体pEGFP-C1。瞬时转染MCF7细胞，Elf5蛋白表达在细胞核中。结论：pEGFP-Elf5真核表达质粒构建成功，Elf5定位在细胞核中。