MK—EGFP融合蛋白在人肝癌细胞Hep—G2中的核定位

目的构建MK—EGFP真核表达质粒，研究该融合蛋白在肝癌细胞株Hep—G2中的定位。方法RT—PCR技术扩增MK编码基因，克隆到真核表达载体pEGFP—N2中，重组质粒pMK—EGFP转染肝癌细胞Hep—c2，显微镜下实时追踪观察融合蛋白的胞内表达与定位。结果成功构建表达质粒pMK—EGFP，真核转染后，在宿主细胞内发生核定位，融合蛋白在核仁区富集。结论转染产生的MK—EGFP融合蛋白在人肝癌细胞Hep—G2中大量表达，并发生明显的核定位现象。

Objective To construct the expressive plasmid of MK - EGFP, and investigate the cellular localization of the fusion protein in hepatocellular cancer cell line HepG - 2. Methods The encoding sequence of MK was amplified by RT - PCR. The eukaryotic expressive plasmid pMK - EGFP was constructed. The recombinant plasmid was transfected into HepG - 2. The expression and cellular localization were observed in real time by confocal laser microscope. Results The recombinant plasmid was identical with expected. Nuclear localization was found in the host cells after transfection. In addition, MK - EGFP was accumulated in the nucleoli. Conclusion The MK - EGFP fusion protein is expressed in HepG - 2 and localized in nuclei.