[myc-p62 fusion protein suppresses the phosphorylation of extracellular signal-regulated kinase].
OBJECTIVE
To construct a eukaryotic expression vector of human autophagy maker protein P62 labeled with myc tag, and detect its biological function.


METHODS
Human P62 gene was amplified from human breast DNA library by PCR and cloned into pXJ-40-myc vector. HEK293T cells were transfected with the recombinant plasmid myc-p62. Western blotting was conducted to detect the fusion protein expression and the effect on the phosphorylation of extracellular signal-regulated kinase (ERK).


RESULTS
Double enzyme identification and sequencing result showed that P62 eukaryotic expression vector labeled with myc tag was successfully constructed and the inserted fragment was correct. Western blotting indicated that the fusion protein was successfully expressed and was able to inhibit ERK phosphorylation.


CONCLUSIONS
The eukaryotic expression vector of myc-p62 was successfully constructed and proved to have an inhibitory effect on ERK phosphorylation.
Second Artillery General Hospital, Beijing 100088, China.
Yunjing
Y
Zhang
Second Artillery General Hospital, Beijing 100088, China.
Yingying
Y
Feng
Division of Health Care, Guard Bureau, PLA General Staff Department, Beijing 100017, China.
Li
L
Zhang
Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Xiaojie
X
Xu
Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Tao
T
Wang
Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Zhongyi
Z
Fan
Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Qinong
Q
Ye
Second Artillery General Hospital, Beijing 100088, China.
Rong
R
Zhang
D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D010766:Phosphorylation; D015202:Protein Engineering; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics* / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*
C489427:MYC protein, human; C118585:P62 protein, human; D016271:Proto-Oncogene Proteins c-myc; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D048049:Extracellular Signal-Regulated MAP Kinases