MK2 different mutants expression and intracellular location of.doc

 PAGE \* MERGEFORMAT 13 MK2 different mutants expression and intracellular location of Author: WEI Jie Gong Xiaoyan Wang Xu Wei-ming, DENG Peng Yong Wang daan [Abstract] Objective: To study the MAPK-activated protein kinase 2 (MAPK-activated protein kinase 2, MK2) cells and their functional relationship between the location to build a MK2 different mutants of green fluorescent protein fusion expression vector and to observe these mutants in the cell positioning within the situation. Methods: The cloning of green fluorescent protein vector pEGFP-C2 on the wild-type MK2 (WT) by site-directed mutagenesis technology to build its non-active mutant of MK2 (320A) and active mutant of MK2 (320E), transfected NIH3T3 cells and in the fluorescence microscope to observe the location of the cell. Results: The variety of MK2 different mutants identified by DNA sequencing is correct, in the NIH3T3 cells have high levels of expression. Send green fluorescent fusion protein showed that in resting state, MK2 (WT) is mainly distributed in the nucleus, MK2 (320E) is mainly distributed in the cytoplasm, and MK2 (320A) in the nucleus and the cytoplasm are distributed. In the under ultraviolet (UV) stimulation, MK2 (WT) and MK2 (320A) shift out of core, while the MK2 (320E) the intracellular location was no obvious change. Conclusion: The successful construction of green fluorescent protein labeled MK2 and the different mutants in eukaryotic cells were expressed in different mutants were observed on the intracellular localization of MK2. [Keywords:] protein kinase; mutation; gene expression; cell positioning; p38 mitogen-activated protein kinase; MAPK-activated protein kinase 2 [Abstract] Objective: To study the intracellular localization of MAPK-activated protein kinase 2 (MAPKAPK2/MK2) and its relation with the function. Methods: Eukaryotic cell expression vectors of green fluorescent protein fused with MAPK-activated protein kinase 2 (MK2) mutants were constructed. Wild-type M