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目的:观察核因子κB(NF-κB)的核定位基序与目的基因质粒重组后,对目的基因转染入核效率的影响。方法:构建含NF-κB核定位基序的人基质细胞衍生因子1α质粒(phSDF-1α-NF-κB),用Cy3核酸荧光染料标记后用聚乙烯亚胺(PEI)转染人脐静脉血管内皮细胞,以不含NF-κB核定位基序的人基质细胞衍生因子1α质粒(phSDF-1α)作对照,比较二者的入核效率和表达效率,以评价NF-κB核定位基序对SDF-1α基因入核和转染的影响。结果:phSDF-1α-NF-κB转染组和phSDF-1α转染组胞核和细胞内Cy3荧光强度比分别为(72±15)%和(12±6)%,二者差异有统计学意义(P<0.01);ELISA结果显示前者SDF-1α蛋白表达量是后者4倍。结论:NF-κB核定位基序能显著促进SDF-1α质粒转染的入核效率,从而提高基因表达水平。
OBJECTIVE: To observe the influence of nuclear localization of nuclear factor κB (NF-κB) on nuclear efficiency after recombination with target gene plasmid. METHODS: Human stromal cell-derived factor-1α plasmid (phSDF-1α-NF-κB) containing NF-κB nuclear localization motif was constructed and transfected into human umbilical vein by polyethyleneimine (PEI) Endothelial cells were transfected with human stromal cell-derived factor 1α plasmid (phSDF-1α) without NF-κB nuclear localization motif as a control, and the efficiency of nuclear import and expression of the two genes were compared to evaluate the localization of NF-κB nuclear localization motif Effect of SDF-1α gene transfection and nuclear transfection. RESULTS: The fluorescence intensity ratios of Cy3 in nuclei and cells in phSDF-1α-NF-κB transfection group and phSDF-1α transfection group were (72 ± 15)% and (12 ± 6)%, respectively, with statistical difference (P <0.01). ELISA results showed that the former SDF-1α protein expression was four times higher than the latter. Conclusion: NF-κB nuclear localization motif can significantly promote the efficiency of nuclear transfection of SDF-1α plasmid transfection, thereby enhancing the gene expression level.