论文部分内容阅读
目的 构建具有绿色荧光蛋白报道 (GFP)基因的端粒酶真核质粒载体 ,为转染细胞、延长细胞寿命提供基础。 方法 酶切含人端粒酶逆转录酶 (h TRT)基因的 p GRN14 5质粒获取 h TRT片段 ,插入经酶切及磷酸化处理的 GFP载体 p EGFP- C1,形成 8.1kb质粒 ,经 Hind 、Not 酶切电泳筛选、鉴定并测序。 结果 经酶切电泳分析得到3.4 kb及 4 .7kb大小的两片段 ;测序结果显示接头两端序列正确。 结论 重组端粒酶 p EGFP- h TRT质粒的成功构建 ,可用于细胞转染 ,对进一步研究组织工程种子细胞的衰老问题具有重要意义。
Objective To construct a plasmid of eukaryotic plasmid with green fluorescent protein reporter gene (GFP), which provides a foundation for cell transfection and cell longevity. Methods The h TRT fragment was obtained by digesting p GRN14 5 plasmid containing human telomerase reverse transcriptase (h TRT) gene and inserted into the pEGFP-C1 GFP vector after digestion and phosphorylation to form the 8.1 kb plasmid. The Hind, Not digestion electrophoresis screening, identification and sequencing. Results The two fragments of 3.4 kb and 4.7 kb were obtained by restriction endonuclease analysis. The sequencing results showed that the two ends of the linker were correct. Conclusion The successful construction of the recombinant telomerase p EGFP-h TRT plasmid can be used for cell transfection and is of great significance for the further study on the senescence of tissue engineering seed cells.