Identifying Of DAPK3:a Interactive Protein Of LOH12CR1

Objective In2011by using the method of the MatchmakerTM Gold Yeast Two-Hybrid System our group screened the possible direct interactive protein of LOH12CR1--DAPK3. LOH12CR1may be a tumor suppressor gene, but its specific biological function and mechanism are unclear. Verifying the real interaction between LOH12CR1and DAPK3in the intracellular may Provide an entry point for the study of LOH12CR1function.Methodsl.We constructed the recombinant vector of pCDNA3.1-myc-his-LOH12CR1and pCMV-tag2-DAPK3,then transfected LOH12CR1and DAPK3into HEK293cells. To verify the interaction between LOH12CR1and DAPK3we used co-immunoprecipitation techniques;2.Confocal laser scanning microscopy is used to observe LOH12CR1’s and DAPK3’s subcellular level positioning in HEK293.Results1.Through coimmunoprecipitation we discovered that LOH12CR1can interact with DAPK3;2.the DAPK3can co-locate with LOH12CR1by coimmunolocaliztion. Conclusion At present promoting apoptosis is the well-known biological function of DAPK3.Our resesrch has confirmed the interaction bewteen LOH12CR1and DAPK3in the intracellular, suggesting that LOH12CR1may be involved in the regulation of apoptotic process through the interaction with DAPK3.We will conduct further studies to clarify the mechanism of it.