摘要
目的观察转染人野生型和突变型早老素1(Presenilin1,PS1)-EGPF后对SH-SY5Y细胞的影响。方法采用定点突变技术构建含突变PS1基因的质粒及与绿色荧光蛋白共表达载体,转染至SY5Y细胞中,筛选出稳定表达的细胞克隆并鉴定;观察转染后各组细胞之间形态的区别,MTT法测定细胞活力并进行比较。结果稳定表达野生型和突变型PS1的细胞模型构建成功,转染pcDNA3.1/PS1突变质粒的细胞活力明显降低。结论此细胞模型的建立为下一步研究突变型PS1在AD发病机制中的作用奠定了基础。
Objective To observe the effect of over-expressing wild and mutative human presenilin I on SH-SYSY cells. Methods We used one step Site-Directed Mutagenesis technique to establish plasmids containing mutative PSI gene, and constructed a co, expressing vector with EGFP. The recombinant plasmid was transfected into SH-SY5Y cells by lipofectamin method and stably transfected cell clones were screened and identified. The differences of cells morphology were observed and cells viability was measured by MTT. Results The SH-SYSY cells model stably expressing the mutative gene were obtained. The cells viability of mutants decreased significantly. Conclusions The establishment of the cells model enable us to explore the biological function of the mutative PSI gene in the pathogenesis of AD.
出处
《卒中与神经疾病》
2006年第6期323-327,共5页
Stroke and Nervous Diseases
基金
国家自然科学基金资助(编号:30370494)
关键词
阿尔茨海默
早老素1
基因突变
绿色荧光蛋白
SH—SY5Y
Alzheimer＇s disease PSl（Presenilin 1） gene mutation EGFP（enhanced green fluorescence protein） SH-SYSY
