[Construction and expression of green fluorescent protein fusion vector of p38 regulated/activated protein kinase].
OBJECTIVE
To construct the expression vector of p38 regulated/activated protein kinase (PRAK) fused to green fluorescent protein which is capable of expression in mammalian cells.


METHODS
PRAK with His-tag in pET-14b expression vector was subcloned into the green fluorescent protein vector pEGFP-C2. The recombinant vector was then transfected into HeLa cells, followed by observation of the cells with fluorescent microscope.


RESULTS
Identification by enzyme digestion, PCR and sequencing confirmed successful construction of the recombinant vector, which was highly expressed in HeLa cells. Green fluorescence of the fusion protein EGFP-PRAK was observed mainly in the cell nuclei.


CONCLUSIONS
The expression vector of PRAK fused to green fluorescent protein is successfully constructed and expressed in mammalian cells, which may facilitate the study of intracellular localization and translocation of PRAK.
Peng
Yi
Y
Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China.
Gong
Xiao-Wei
XW
Deng
Peng
P
Qin
Qing-He
QH
Jiang
Yong
Y
D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000096:biosynthesis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D048051:p38 Mitogen-Activated Protein Kinases