dRASSF在果蝇细胞中负调控Dmp53的蛋白水平

dRASSF Negatively Regulates Dmp53 in Drosophila Cells

从果蝇细胞cDNA文库克隆得到基因Dmp53,将该基因插入到pAc-HisB载体中,得到的重组DNA成功在果蝇S2细胞中表达.通过免疫荧光对dRASSF和Dmp53亚细胞定位进行了分析,确定了两者均是典型细胞核定位且共转染时存在共定位现象.免疫共沉淀和western blotting分析未检测到dRASSF和Dmp53之间存在相互结合作用,但是dRASSF过表达可以有效地降低Dmp53的蛋白量.为深入阐释dRASSF和Dmp53间的相互关系提供有益的线索.

The Dmp53 gene was amplified from the cDNA library of Drosophila S2 cells and inserted into vector pAc-HisB.The recombination DNA was then successfully expressed in S2 cells.The fluorescence assay indicated that dRASSF and Dmp53 predominately localize in the nucleus and colocalize when their co-expressing in S2 cells.Immunoprecipitation and western blotting analysis suggested that overexpression of dRASSF results in downregulation of Dmp53 protein level,although there is no detectable physical interaction between dRASSF and Dmp53.This study support further investigation about the mechanism of how dRASSF regulates Dmp53.