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《Journal of Xi'an Jiaotong University(Medical Sciences)》 2016-02
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The eukaryotic expression,intracellular location and functions of human CAP1

LIU Xu;ZHANG Ying;WANG Bin;LIU Xiao-dan;WANG Yu;ZENG Yan;PAN Xiu-jie;ZHOU Ping-kun;ZHU Mao-xiang;GU Yong-qing;College of Life Sciences,Shihezi University;Institute of Radiology and Radiation Medicine,the Chinese Academy of Military Medicine;College of Medicine,Shihezi University;  
Objective To construct the recombinant eukaryotic expression plasmids of human adenylyl cyclase-associated protein1(CAP1)and to explore its intracellular location and functions.Methods By using Hela cDNA as the template,the cDNAs encoding CAP1 was amplified by PCR and inserted into pCMV-Myc vector to construct the recombinant plasmid.The recombinant plasmid was transfected into 293 cells using lipofectamine2000.The protein expression and the intracellular location of the inserted gene were confirmed by Western blotting and immunofluorescence,respectively.Scratch-repair experiment was used to detect the cancer cells'migration ability.Results The recombinant eukaryotic expression plasmid of human CAP1 was successfully constructed and transfected into eukaryote cells.The recombinant plasmid was successfully expressed in eukaryote cells.CAP1 was located in the cytoplasm.The results of scratch-repair experiment showed that the overexpression of CAP1 could significantly inhibit the cells'migration.Conclusion CAP1 recombinant plasmid was successfully expressed in eukaryotic cells.CAP1 protein was located in the cytoplasm.The overexpression of CAP1 inhibited cell migration.The present study provides important experimental evidence for further study on CAP1.
【Fund】: 国家自然科学基金资助项目(No.30960093 31160183 31270894 31470827)~~
【CateGory Index】: R3416
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