Construction and characterization of cardiac specific promoter-driven expression vector / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1546-1551, 2009.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-296892
Responsible library:
WPRO
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.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Transfection
/
Cell Differentiation
/
Promoter Regions, Genetic
/
Electroporation
/
DNA, Complementary
/
Myosin Heavy Chains
/
Cell Biology
/
Cytomegalovirus
/
Green Fluorescent Proteins
/
Embryonic Stem Cells
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2009
Document type:
Article