title: A dual color fluorescent reporter system for the real time detection of promoter activity.
abstract: Understanding the mechanisms controlling transcription of a gene requires the identification and characterization of its cis-acting regulatory elements. A highly useful approach to the identification and characterization of cis-acting elements has been the systematic coupling of genomic fragments to reporter constructs, so called "promoter bashing". The expression from such reporters must be normalized for differences in transient transfection efficiency between cells and replicates. A novel dual color fluorescent reporter system to assay the promoter activity of a genomic DNA fragment of interest was established by cloning a Discosoma red fluorescent protein gene and a green fluorescent protein gene into a single vector, giving a system in which the ratio between red and green fluorescence is proportional to promoter activity. This system allows real time quantitative monitoring of promoter activity. We validated this approach by assaying the cis-acting regulatory potential of the peroxisome proliferators-activated receptor gamma2 gene.
authors.affiliation: College of Animal Science and Technology, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, No. 22 Xinong Road, 712100, Yangling, Shaanxi, China.
authors.forename: Liushuai
authors.initials: L
authors.lastname: Hua
authors.forename: Mingxun
authors.initials: M
authors.lastname: Li
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Sun
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Zhuanjian
authors.initials: Z
authors.lastname: Li
authors.forename: Yao
authors.initials: Y
authors.lastname: Xu
authors.forename: Shenrong
authors.initials: S
authors.lastname: Hu
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D037421:Anthozoa; D003001:Cloning, Molecular; D003116:Color; D005456:Fluorescent Dyes / Q000378:metabolism*; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D013194:Staining and Labeling / Q000379:methods*; D014158:Transcription, Genetic; D000097573:Red Fluorescent Protein
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein