[Construction and expression of red fluorescent protein fusion vector incorporating p38 mitogen-activated protein kinase].
OBJECTIVE
To construct the vector that expresses the fusion protein of p38 mitogen-activated protein kinase (MAPK) and red fluorescent protein (RFP) in mammalian cells.


METHODS
FLAG-tagged p38 MAPK in pcDNA3 vector was subcloned into RFP vector pDsRed1-N1, the construct of which was then transfected into HeLa cells and observed with fluorescence microscope.


RESULTS
The recombinant plasmid was verified by enzyme digestion, PCR and sequence analysis, and p38 MAPK-RFP fusion protein was highly expressed in HeLa cells. Fluorescence microscope found the red fluorescence distributed all over the cytoplasm and in the nuclei as well.


CONCLUSIONS
The expression vector for p38 MAPK-RFP fusion protein is successfully constructed and effective expression of this fusion protein is achieved, which might be instrumental in the study of intracellular localization of p38 MAPK.
Gong
Xiao-Wei
XW
Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Qin
Qing-He
QH
Wang
Jing-Zhen
JZ
Jiang
Yong
Y
D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D020928:Mitogen-Activated Protein Kinases / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D020928:Mitogen-Activated Protein Kinases; D011993:Recombinant Fusion Proteins; D048051:p38 Mitogen-Activated Protein Kinases; D000097573:Red Fluorescent Protein