PC-1分子在细胞内定位的深入探讨

Further Study on The Localization of PC-1

为了寻找影响PC 1分子进入细胞核的序列 ,将该分子不同区段分别与EGFP融合表达 ,通过观察绿色荧光蛋白在细胞内的分布 ,发现PC 1分子的第 112～ 190区段是一独立的功能区 ,缺失这一区段后 ,分子的其余部分能够进入细胞核并在其中积累 ,这一区段使PC 1分子被排斥于细胞核之外 ,并在细胞质中浓缩成许多点状结构 .运用以SOS恢复系统为基础的酵母双杂交方法 ,发现PC 1分子能够定位于细胞膜上 ,通过缺失突变发现该分子的第 112～ 190区段是一独立的细胞膜定位信号序列 ,这一序列将其定位于细胞的胞膜上 .

In order to search for the sequence that affects the entering of PC-1 into cell nucleus,different part of PC-1 cDNA were fused to that of EGFP. Through the observation of the distribution of green fluorescence in the cell,the region between 112～190 amino acids was identified as independent functional domain which excluded PC-1 from cell nucleus and there appeared many dotted structure in the cell cytoplasm. The other part of PC-1 could accumulated in the nucleus when this region was deleted. Simultaneously,PC-1 was also found to attach to the cell membrane by the yeast two-hybrid system based on the SOS recovery sytem,what is more,the region between 112～190 amino acids was identified as independent domain responsible for it is cell membrane anchoring.