2011•Tianjin yiyaoRequires access
Construction and Expression of pEGFP-C2-hG3BP-Domain (1-5) Recombinant Eukaryotic Plasmids
Mengyu Zhu, Xingjie Gao, Baoxin Qian, Ge Lin, Hong Zhao, Zhao Xiujuan, Jie Yang
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Abstract
Objective:To construct eukaryotic green fluorescent protein (GFP) expressing recombinant plasmids,pEGFP-C2-hG3BP-Domain (1-5),which contain domain (1-5) fragments of human G3BP.Methods:The genes of G3BP fragments were amplified by PCR from the recombinant pEGFP-C1-G3BP plasmid and inserted into pEGFP-C2 fluorescent expressing vector with EcoRI and BamHI sites.These recombinant pEGFP-C2-hG3BP-Domain(1-5) plasmids were transfected into HeLa cells and the expression of green fluorescent fusion proteins was examined by fluorescence microscope and Western blotting assay.Results:The domain (1-5) fragments of G3BP were sequenced correctly and detected in the products of the restriction single/double enzyme digestion.The green fluorescent fusion proteins were also detected in the transfected HeLa cells by fluorescence microscope and Western blotting assay.Conclusion:These recombinant eukaryotic plasmids of pEGFP-C2-G3BP-Domain (1-5) were constructed successfully and expressed effectively.