国家自然科学基金资助项目(30070296)和国家高技术“863”计划资助项目(2002AA223061).
This work was supported by grants from The National Natural Science Foundation of China(30070296) and State 863 High Technology R&D Project of China(2002AA223061).
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.
张浩,周建光,李杰之,黄翠芬. PC-1分子在细胞内定位的深入探讨[J].生物化学与生物物理进展,2004,31(3):260-266
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