[Construction and characterization of cardiac specific promoter-driven expression vector].
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.
He
Wenjun
W
Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Li
Shichong
S
Ye
Lingling
L
Wang
Qiwei
Q
Wang
Haitao
H
Xie
Jing
J
Liu
Hong
H
Chen
Zhaolie
Z
D000818:Animals; D002454:Cell Differentiation; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D018274:Electroporation; D053595:Embryonic Stem Cells / Q000166:cytology*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D018995:Myosin Heavy Chains