[Construction and identification of recombinant firefly luciferase report vector containing human acyl coenzyme a: cholesterol acyltransferase 1 gene P7 promoter].
The DNA segment of the human acyl coenzyme A: cholesterol acyltransferasel (ACAT1) gene P7 promoter was amplified by PCR from human monocytic leukemia cell line (THP-1) and cloned to TA vector, then the positive clone was confirmed by restriction enzymes and sequencing. The targeted segment was subcloned to Firefly luciferase report vector pGL3-Enhancer. The recombinant plasmid pGL3E-P7 was transfected transiently into THP-1, then the expression of luciferase could be detected in THP-1 by pGL3E-P7 transfection. We successfully constructed luciferase reporter vector containing P7 promoter of the human ACAT1 gene, and established a new means to study the transcriptional regulation mechanisms of ACAT1 during atherosclerosis.
Ge
Jing
J
Department of Gerontology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China.
Cheng
Bei
B
He
Ping
P
Wen
Hui
H
Lu
Han
H
Chen
Xin
X
Zeng
Yongli
Y
D045744:Cell Line, Tumor; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007948:Leukemia, Monocytic, Acute / Q000473:pathology; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D002785:Sterol O-Acyltransferase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D012333:RNA, Messenger; D011994:Recombinant Proteins; D049409:Luciferases, Firefly; D002785:Sterol O-Acyltransferase; C493492:sterol O-acyltransferase 1