Related Experiment Videos
[Construction and expression of red fluorescent protein fusion vector incorporating p38 mitogen-activated protein
Xiao-Wei Gong1, Qing-He Qin, Jing-Zhen Wang
1Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
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.
Conclusion:
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.