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《Chinese Journal of Clinical Medicine》 2018-04
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Effect of myc/his-TDP-43 fusion protein on alternative splicing of tau exon 10

MIAO Shi-chen;QU Shu-ting;LU Ying;TANG Xiao;REN Liang-liang;GU Jian-lan;School of Medicine,Nantong University;  
Objective:To construct plasmids of TDP-43 and its truncations to pcDNA3.1 vector with myc/his tag and explore the effect of myc/his-TDP-43 fusion protein on the alternative splicing of tau exon 10.Methods:The cDNA of TDP-43 and its truncations were amplified by polymerase chain reaction(PCR)and inserted into pcDNA3.1 after endonuclease cleavage.The recombinant plasmids were transfected into HEK-293 FT cells and the expression of TDP-43 were detected by Western blots.TDP-43 or siTDP-43 was co-transfected with tau mini gene,SI9/SI10,and the alternative splicing products of tau exon 10 was detected by RT-PCR.Results:pcDNA3.1/TDP-43·myc·his and the truncation plasmids were constructed successfully.Myc/his-TDP-43 and truncation fusion proteins were expressed in HEK-293 FT cells.Myc/his-TDP-43 fusion protein induced the expression of 4 R-tau.Conclusions: Myc/his-TDP-43 fusion protein promoted 4 R-tau expression.The truncation plasmids of TDP-43 might be helpful for the future study of the function of different domains of TDP-43 in tau pathology and alternative splicing of tau exon 10 in neurodegenerative diseases.
【Fund】: 大学生创新创业计划国家级项目(201710304031Z);; 江苏省高校自然科学基金面上项目(16KJB310013);; 南通市市级科技计划项目(MS12015058);; 南通大学博士科研启动基金(15B30)~~
【CateGory Index】: R749.16
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