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目的:研究人野生型DNA聚合酶β(wtDNApolβ)在NIH3T3细胞内的定位。方法:将携带人wtDNApolβ的真核绿色荧光表达载体pEGFP-C3-polβ,用脂质体介导的方法转染NIH3T3细胞株,用荧光倒置显微镜观察转染细胞,观察wtpEGFP-C3-polβ融合基因表达。以转染空质粒pEGFP-C3的NIH3T3细胞为对照。结果:转染细胞均有GFP表达,说明转染成功。转染pEGFP-C3-polβ的细胞胞核荧光强度较胞浆强,而空质粒转染细胞胞浆胞核荧光强度无差别。结论:wtDNApolβ在NIH3T3细胞中定位于胞核内。
Objective: To study the localization of human wild-type DNA polymerase β (wtDNApolβ) in NIH3T3 cells. Methods: The eukaryotic expression vector pEGFP-C3-polβ carrying human wtDNApolβ was transfected into NIH3T3 cells by liposome-mediated method. The transfected cells were observed by fluorescence inverted microscope and the wtpEGFP-C3-polβ fusion gene expression. The NIH3T3 cells transfected with empty plasmid pEGFP-C3 were used as control. Results: The transfected cells had GFP expression, indicating that the transfection was successful. The fluorescence intensity of cells transfected with pEGFP-C3-polβ was stronger than that of the cytoplasm, while the fluorescence intensity of cytoplasmic nucleus of empty plasmid transfected cells had no difference. Conclusion: wtDNApolβ is localized in nucleus in NIH3T3 cells.