Construction and expression of green fluorescent protein fusion vector of p38 regulated/activated protein kinase

Abstract

Abstract: 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.Conclusion 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.

CLC Number:

PENG Yi, GONG Xiao-wei, DENG Peng, QIN Qing-he, JIANG Yong. Construction and expression of green fluorescent protein fusion vector of p38 regulated/activated protein kinase[J]. Journal of Southern Medical University, 2005, 25(03): 285-288.

References

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