含p38丝裂原活化蛋白激酶的红色荧光蛋白融合载体的构建与表达
[Construction and expression of red fluorescent protein fusion vector incorporating p38 mitogen-activated protein kinase].
作者信息
Gong Xiao-Wei, Qin Qing-He, Wang Jing-Zhen, Jiang Yong
机构信息
Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
出版信息
Di Yi Jun Yi Da Xue Xue Bao. 2002 Feb;22(2):171-3.
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.
目的
构建能在哺乳动物细胞中表达p38丝裂原活化蛋白激酶（MAPK）与红色荧光蛋白（RFP）融合蛋白的载体。
方法
将pcDNA3载体中带有FLAG标签的p38 MAPK亚克隆至RFP载体pDsRed1-N1中，构建好的载体随后转染至HeLa细胞中，并用荧光显微镜观察。
结果
重组质粒经酶切、PCR及序列分析验证，p38 MAPK-RFP融合蛋白在HeLa细胞中高表达。荧光显微镜观察发现红色荧光遍布细胞质及细胞核。
结论
成功构建了p38 MAPK-RFP融合蛋白的表达载体，并实现了该融合蛋白的有效表达，这可能有助于研究p38 MAPK的细胞内定位。
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