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Construction and expression of the pIRES-enchanced green fluorescent protein reporter gene vector carrying BMP_2 Gene

ZOU Haibo,et al (Department of Orthopaedic Surgery,College of Clinical Medicine,Chongqing Medical University)  
Objective:To construct the pIRES2-enchanced green flurescent protein expression vector carrying BMP_2 gene in order to provide an ideal reporter gene for the expression and identify the location of portein in vitro and vivo.Methods:The BMP_2 cDNA sequence was excised,and subcloned on pUC18 vector.And then the BMP_2 sequence on the pUC18 plasmid was excised again and ligated into Sal Ⅰ site of pIRES2-EGFP treated by phosphorylase in sense and antisense direction.The correct recombinant plasmid was transfected into 293-T cells,the expression of BMP_2 was determined by FCM,LCM and RT-PCR.Results:The recombinant expression vector was verified correctly by enzyme digestion,sequence analysis.This vector was expressed in cells.Conclusion:A pIRES2 green fluorescent protein reporter gene vector containing BMP_2 has been constructed successfully,thus providing an important and convenient tool to study intracellular localization of BMP_2.
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