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| Construction of psiCHECK-2eGFP-mLumin dual fluorescent protein reporter gene and expression in eukaryotic cells |
| PAN Xiao-yuan, WU Wei-jiang, LIU Yun, ZHANG Miao-miao, LIU Xia,DU Feng-yi, XIA Sheng, SHAO Qi-xiang |
| （School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang Jiangsu 212013, China） |
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Abstract Objective: To construct a new dual fluorescent protein reporter gene plasmid, psiCHECK-2-eGFP-mLumin. Methods: The dual fluorescent protein reporters gene plasmid was constructed by inserting the PCR amplification products of enhanced green fluorescent protein (eGFP) and mLumin into psiCHECK-2 vector to replace the firefly luciferase and renilla luciferase, respectively. The sequence of eGFP and mLumin were confirmed by PCR, and blasted online of the National Center for Biotechnology Information. HEK293T cells were transfected with psiCHECK-2-eGFP-mLumin plasmid and the expression efficiency of the fluorescent proteins was valuated. Results: The dual fluorescent protein reporters gene plasmid, psiCHECK-2-eGFP-mLumin, was successfully constructed by inserting the correct sequence of eGFP and mLumin into the psiCHECK-2 plasmid. The psiCHECK-2-eGFP-mLumin successfully mediated the gene expression of eGFP and mLumin in HEK293T cells. Conclusion: The psiCHECK-2-enhanced green fluorescent protein-mLumin plasmid was constructed successfully and expressed well in HEK293T cells.
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Received: 28 May 2013
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2013, Vol. 23 
