Tat-p21融合蛋白的表达及其在A549细胞中的活性研究.doc

文档介绍：
Tat-p21 融合蛋白的表达及其在 A549 细胞
中的活性研究#
施惠敏,吴兰,赵梓亦,吴传芳**
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(四川大学生命科学学院)
摘要:p21 蛋白是细胞周期依赖性蛋白激酶的抑制剂,它可以和许多细胞周期蛋白/细胞周期
蛋白激酶复合物结合,使受到 DNA 损伤的细胞停滞于 G1 p21 蛋白可以
Tat 广泛应用于目的蛋白转导进入细胞,使被
,将 p21 蛋白与 Tat 融合,按浓度梯度:
100,200,400,800,1000nmol/L 与 A549 细胞共同孵育, 确定出最佳浓度, 并分别于
10,60,120,240min 终止反应,以找到最佳的孵育时间;接着检测了转导进入 A549 细胞的
Tat-p21 融合蛋白在细胞中稳定存在的时间;并通过绘制细胞生长曲线的方法比较了转导
Tat-p21 的细胞与负对照实验组生长速度的区别.。
关键词: Tat;p21 蛋白;A549 细胞
中图分类号:Q291
Expression of Tat-p21 Fusion Protein and Its Activity in
A549 Cells
shihuimin , wulan , zhaoziyi , wuchuanfang
sichuan university. coliiege of life science,chengdu,610064;
Abstract: p21 is a potent cyclin-dependent kinase inhibitor (CKI). The p21 (CIP1/WAF1) protein
binds to and inhibits the activity of a number of cyclin and plexes, which imposes cell
cycle block in G1 and enables the DNA repair program of restoring genomic integrity. A direct
modulation of p21 protein could affect cell proliferation. binant protein fused to protein
transduction domain(PTD) of TAT protein(TAT-PTD) is widely utilized to deliver protein of
biological function to a variety of cell types. In this study, a binant protein Tat-p21 was
constructed and expressed in vitro and incubated with A549 cells in a concentration series of
100,200,400,800,1000nmol/L. Each group of cells was harve