Rapid and direct quantitative RT-PCR method to measure promoter activity.
This Note describes a novel rapid and direct quantitative method for examining the activity of genetic response elements. This method will provide an alternative to the classically used "reporter gene" activity assays. We show that a transfected genetic cis-regulatory element that responds to the transcription factor p53 gives a quantitative read-out at the RNA level that parallels that of an endogenous p53 responsive gene, p21 waf1/cip1. The correlation between the endogenous p21 gene expression in response to p53 and the transfected cis element is remarkable. This method is more direct and potentially faster than traditional promoter-reporter assays.
Yu
Lei
L
Free Radical and Radiation Program, Radiation Oncology Department, B180 ML, The University of Iowa, Iowa City, Iowa 52242, USA.
Domann
Frederick E
FE
D052160:Benzothiazoles; D001709:Biotechnology / Q000379:methods*; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000737:chemistry; D003959:Diamines; D017930:Genes, Reporter; D016158:Genes, p53; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D009930:Organic Chemicals / Q000494:pharmacology; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011804:Quinolines; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000502:physiology
D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins; C098022:SYBR Green I; D012313:RNA