Construction and identification of recombinant firefly luciferase report vector containing human acyl coenzyme a: cholesterol acyltransferase 1 gene P7 promoter.
- Author:
Jing GE
1
;
Bei CHENG
;
Ping HE
;
Hui WEN
;
Han LU
;
Xin CHEN
;
Yongli ZENG
Author Information
1. Department of Gerontology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Chromosomes, Human, Pair 7;
genetics;
Gene Expression Regulation;
Genes, Reporter;
genetics;
Genetic Vectors;
genetics;
Humans;
Leukemia, Monocytic, Acute;
pathology;
Luciferases, Firefly;
genetics;
metabolism;
RNA, Messenger;
genetics;
metabolism;
Recombinant Proteins;
genetics;
metabolism;
Sterol O-Acyltransferase;
genetics;
metabolism;
Transfection
- From:
Journal of Biomedical Engineering
2008;25(6):1381-1384
- CountryChina
- Language:Chinese
-
Abstract:
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.
