Construction of eukaryotic expression plasmid of human PRX3 and its expression in HEK-293FT cells.
To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Feng
Yan
Y
Department of Pathology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Liu
Zhao
Z
Cao
Huiqing
H
Meng
Xianmin
X
Qu
Zhiling
Z
Xiong
Mi
M
Deng
Zhongduan
Z
D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004622:Embryo, Mammalian; D005057:Eukaryotic Cells / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D010544:Peroxidases / Q000096:biosynthesis* / Q000235:genetics; D061105:Peroxiredoxin III; D054464:Peroxiredoxins; D010957:Plasmids / Q000235:genetics*; D014162:Transfection
D010544:Peroxidases; C516115:PRDX3 protein, human; D061105:Peroxiredoxin III; D054464:Peroxiredoxins