论文部分内容阅读
目的以绿色荧光蛋白（ＧＦＰ）作为一个标记，观察ＧＦＰ标记的ＨＴＨ１基因修饰的ＮＩＨ－３Ｔ３细胞在体外及植入脑内后的生长情况，判断ＧＦＰ基因能否作为一种新的筛选标记基因。方法构建同时表达ＧＦＰ和ＨＴＨ１双基因的多基因表达载体ＧＣ－ＧＦＰ－Ｐ－ＨＴＨ１－ＳＮ。并转染ＮＩＨ－３Ｔ３细胞，且移植入Ｐａｒｋｉｎｓｏｎ病模型大鼠纹状体内。结果用荧光显微镜、流式细胞仪、免疫组化、ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ等方法均检测到ＧＦＰ、ＨＴＨ１在体外和脑内的表达。结论ＧＦＰ可作为一个筛选标记基因，可对目的基因或植入细胞的其他功能性产物的表达进行观察和检测。
Objective To observe the growth of GFP-labeled HTH1 gene-modified NIH-3T3 cells in vitro and after implantation into the brain with green fluorescent protein (GFP) as a marker to determine whether GFP gene can be used as a new screening marker gene. Methods Constructed multi-gene expression vector GC-GFP-P-HTH1-SN which expressed both GFP and HTH1 double genes simultaneously. And transfected into NIH-3T3 cells, and transplanted into Parkinson’s disease model rat striatum. Results The expression of GFP and HTH1 in vitro and in vivo were detected by fluorescence microscopy, flow cytometry, immunohistochemistry and Western blotting. Conclusion GFP can be used as a screening marker gene to observe and detect the expression of target genes or other functional products implanted in cells.