细胞内活化的Notch途径报告基因质粒的构建和鉴定

Construction and identification of a reporter plasmid for analyses of Notch signaling pathway

目的构建并鉴定一种检测细胞内活化的Notch信号途径的报告基因质粒,用于研究该途径在宫颈癌细胞中的作用机制。方法构建报告基因质粒pTP1-EGFP-N1;测序及酶切鉴定后,与Notch的胞内段(NIC)共转染宫颈癌Hela细胞,观察其表达。结果构建了报告基因质粒pTP1-EGFP-N1,与NIC共转染Hela细胞后有阳性表达。结论该报告基因质粒的成功构建为进一步研究Notch信号途径在宫颈癌发生中的作用机制奠定了实验基础。

Objective To construct and identify a reporter plasmid for analyses of the mechanism of Notch signaling pathway in human cervical carcinoma cells. Methods The reporter gene plasmid pTP1-EGFP-N1 was constructed. It was confirmed by restriction enzyme digestion and DNA sequencing. To observe its expression in Hela cells by co-transfecting the reporter gene plasmid and the intracellular domain of Notch （NIC）. Results Reporter gene plasmid pTP1-EGFP-N1 was successfully constructed. While it was co-transfected into Hela cells with NIC, EGFP was expressed. Conclusion Construction of pTP1-EGFP-N1 will provide a basis for a further study of the mechanism of Notch signaling pathway in the cervical carcinoma.