中文摘要：

目的构建并鉴定转录因子Kaiso的表达质粒pEGFP-Kaiso。方法采用PCR的方法扩增pCS2-Kaiso中人Kaiso的全长序列,克隆至载体pEGFP-N1中,构建重组质粒pEGFP-Kaiso,以酶切及基因测序方法鉴定重组质粒的准确性。利用LipofectamineTM2000将重组质粒转染至肺癌细胞系A549,24 h后荧光显微镜观察绿色荧光信号,48 h后Western blot鉴定Kaiso蛋白表达变化。结果限制性双酶切及DNA测序分析结果显示插入的DNA片段长度约为2 022 bp,与预期Kaiso基因片段大小相同。转染pEGFP-Kaiso重组质粒后,荧光显微镜观察到明确的绿色荧光蛋白（GFP）的绿色荧光信号,Western blot证实转染pEGFP-Kaiso质粒后的Kaiso蛋白表达量显著上调（P〈0.05）。结论成功构建了人Kaiso基因真核表达载体,为深入研究Kaiso的生物学功能提供了有力的实验工具。

英文摘要：

Objective To study the biological function of transcription factor Kaiso in the lung cancer cells and construct the eukaryotic expression vector of human Kaiso gene.Methods Kaiso cDNA was amplified by PCR and inserted into eukaryotic expression vector pEGFP.The construction of the recombinant pEGFP-Kaiso was tested by restriction enzyme analysis and gene sequencing.Then the pEGFP-Kaiso was transfected into lung cancer cell lines,A549 cells.Last,the localization and expression of Kaiso in transfected A549 cells were verified by fluorescent microscope and Western blot,respectively.Results The size of inserted fragments in the recombinant vectors pEGFP-Kaiso is 2 022 bp,corresponding to that of Kaiso.After transfecting pEGFP-Kaiso into A549 cells by using LipofectamineTM 2000,The GFP signal was detected in the nucleus and cytoplasm,while the expression level of Kaiso in transfected A549 cells was significantly elevated（P 0.05）.Conclusion The eukaryotic expression vector pEGFP-Kaiso is successfully constructed and it provides a powerful tool for studying the biological function of transcription factor Kaiso.