[Construction and expression of red fluorescent protein fusion vector incorporating p38 mitogen-activated protein kinase].
Xiaowei Gong, Qing-he Qin, Jingzhen Wang, Yong Jiang
Abstract
Xiaowei Gong, Qing-he Qin, Jingzhen Wang, Yong Jiang
Abstract
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.
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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.
Key concepts: Fusion protein, Protein kinase A, Green fluorescent protein, Molecular biology, Expression vector, Transfection, Fluorescence microscope, HeLa