Title: Construction and eukaryon expression of plRES2-EGFP-CB_2 plasmid
Abstract: Objective:To construct the pIRES2-EGFP-CB2 plasmid and investigate its expression in HEK293 cells.Methods:Full length DNA of CB2 receptor gene amplified from plasmid pcDNA3.1(+)-CB2 using PCR was inserted into pIRES2-EGFP plasmid by gene recombinantion technology.The recombinant plasmid was transfected into HEK293 cells.The expression of EGFP was examined under fluorescence microscope and the expression of CB2 was examined by RT-PCR and Reporter gene.Results:The specificity of PCR products by DNA scquencing and restriction endonuclease reactions demonstrated that recombinant pIRES2-EGFP-CB2 plasmid was successfully constructed.Green fluorescence expression of the 293 cells can be observed under fluorescence microscope and high expression of CB2 was detected in HEK293 cells by RT-PCR and dual luciferase reporter gene;the CB2 receptor agonist JWH-015 could significantly reverse the luciferase activity enhancement which induced by forskolin.Conclusion:pIRES2-EGFP-CB2 has been successfully cloned,which provided a basis for further investigation of CB2's physiological and physiopathological functions and regulatory mechanism.
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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