融合蛋白_____结构域；基因；融合蛋白；纯化

GSThPBD融合蛋白的克隆、表达及活化Rac1蛋白的分析

http://www.100md.com 《第四军医大学学报》 2004年第13期

融合蛋白,,结构域；基因；融合蛋白；纯化,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Cloning and expression of GSThPBD fusion protein for activated Rac1 protein analysis

    HAN YaLin， MENG ZiMin， KANG Jian， LIU HaiWei， YAN ChengHui， WANG ShiWen

    1Department of Cardiovasology, General Hospital of Chinese PLA, Shenyang Command, Shenyang 110016, China, 2Institute of Cardiovascular Diseases, General Hospital of Chinese PLA, Beijing 100850, China

    【Abstract】 AIM: To induce and express the GSThPBD fusion protein and to study the expression of Rac1 in endothelial cell line ECV304. METHODS: The gene fragment of hPBD was amplified by RTPCR and cloned into the pGEX2T prokaryotic expression vector, and the GSThPBD fusion protein was purified. The activated Rac1 protein was detected with GSThPBD by pull down assay. RESULTS: DNA sequencing confirmed that the expression vector of pGEX2T/hPBD was constructed successfully and fusion protein was expressed effectively in Escherichia coli, with about 75% of the GSThPBD fusion protein (Mr 36 000). Pull down assay found the activated Rac1 protein in ECV304 cells. CONCLUSION: The pGEX2T prokaryotic expression vector is successfully constructed and the GSThPBD fusion protein is purified. Activated Rac1 protein is detected in vascular endothelial cell ECV304. This study lays the basis for the research on the relationship between the Rac1 activity and vascular endothelial cell permeability. ......

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