Construction and characterization of cardiac specific promoter-driven expression vector.
- Author:
Wenjun HE
1
;
Shichong LI
;
Lingling YE
;
Qiwei WANG
;
Haitao WANG
;
Jing XIE
;
Hong LIU
;
Zhaolie CHEN
Author Information
1. Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Differentiation;
Cytomegalovirus;
genetics;
metabolism;
DNA, Complementary;
genetics;
Electroporation;
Embryonic Stem Cells;
cytology;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Mice;
Myocardium;
cytology;
metabolism;
Myosin Heavy Chains;
biosynthesis;
genetics;
Promoter Regions, Genetic;
genetics;
Transfection
- From:
Chinese Journal of Biotechnology
2009;25(10):1546-1551
- CountryChina
- Language:Chinese
-
Abstract:
We constructed and identified cardiac-specific a-myosin heavy chain (alpha-MHC) promoter-driven expression vector. alpha-MHC promoter was amplified by PCR by using mouse genomic DNA as template and inserted into pGEM-T Easy vector. The inserted fragment was released by enzyme digestion, and then the cytomegalovirus (CMV) promoter in pcDNA3.1(+)-EGFP-hygro vector was replaced by the alpha-MHC promoter to construct alpha-MHC-EGFP expression vector. After identification with enzyme digestion, alpha-MHC-EGFP was transfected into mouse primary cardiomyocytes by electroporation. Green fluorescence could be observed in transfected cardiomyocytes, but not in transfected non-cardiomyocytes. Alpha-MHC-EGFP expression vector was specifically expressed in cardiomyocytes, and could be used to purify embryonic stem cell-derived cardiomyocytes.
