Construction and expression of the pIRES-enchanced green fluorescent protein reporter gene vector carrying BMP_2 Gene
- VernacularTitle:携带IRES的BMP_2绿色荧光蛋白表达载体的构建和表达
- Author:
Haibo ZOU
- Publication Type:Journal Article
- Keywords:
BMP2;
Green fluorescent protein;
Internal ribosomes entry site;
Eukaryotic expression
- From:
Journal of Chongqing Medical University
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
