BIOMAT Database Logo BIOMAT Database Logo

Development of dual-visible reporter assays to determine the DNA-protein interaction. 2023

Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Key Laboratory of National Forestry and Grassland Administration/Beijing for Bamboo and Rattan Science and Technology, Beijing, 100102, China.

The application of DNA-protein interaction reporter assays for relational or ratiometric measurements within an experimental system is popular in biological research. However, the existing reporter-based interaction assays always require special equipment, expensive chemicals, and a complicated operation. Here, we developed a DNA-protein interaction technology integrating two visible reporters, RUBY and UV-visible GFP (eYGFPuv), which allows the expression of the cassette reporter contained cis-acting DNA element (DE) fused upstream of TATA box and RUBY, and a constitutive promoter regulating eYGFPuv in the same construct. The interaction of transcription factor (TF) and the DE can be detected by co-expressed the cassette reporter and TF in tobacco leaves where the cassette reporter alone serves as a control. We also revealed that eight function-unknown bamboo AP2/ERFs interacted with the DE of ANT-AP2R1R2 (ABE), DRE (DBE), GCC-box (EBE), and RAV1 binding element (RBE), respectively, which are consistent with the results by dual-luciferase reporter assays. Thus, the dual-visible reporters offer a convenient, visible, and cost-saving alternative to other existing techniques for DNA-protein interaction in plants.

UI MeSH Term Description Entries
D011401 Promoter Regions, Genetic DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes. rRNA Promoter,Early Promoters, Genetic,Late Promoters, Genetic,Middle Promoters, Genetic,Promoter Regions,Promoter, Genetic,Promotor Regions,Promotor, Genetic,Pseudopromoter, Genetic,Early Promoter, Genetic,Genetic Late Promoter,Genetic Middle Promoters,Genetic Promoter,Genetic Promoter Region,Genetic Promoter Regions,Genetic Promoters,Genetic Promotor,Genetic Promotors,Genetic Pseudopromoter,Genetic Pseudopromoters,Late Promoter, Genetic,Middle Promoter, Genetic,Promoter Region,Promoter Region, Genetic,Promoter, Genetic Early,Promoter, rRNA,Promoters, Genetic,Promoters, Genetic Middle,Promoters, rRNA,Promotor Region,Promotors, Genetic,Pseudopromoters, Genetic,Region, Genetic Promoter,Region, Promoter,Region, Promotor,Regions, Genetic Promoter,Regions, Promoter,Regions, Promotor,rRNA Promoters
D012045 Regulatory Sequences, Nucleic Acid Nucleic acid sequences involved in regulating the expression of genes. Nucleic Acid Regulatory Sequences,Regulatory Regions, Nucleic Acid (Genetics),Region, Regulatory,Regions, Regulatory,Regulator Regions, Nucleic Acid,Regulatory Region,Regulatory Regions
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D005786 Gene Expression Regulation Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation. Gene Action Regulation,Regulation of Gene Expression,Expression Regulation, Gene,Regulation, Gene Action,Regulation, Gene Expression
D014157 Transcription Factors Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process. Transcription Factor,Factor, Transcription,Factors, Transcription
D017930 Genes, Reporter Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest. Reporter Genes,Gene, Reporter,Reporter Gene

Related Publications

Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Development of reporter gene assays to determine the bioactivity of biopharmaceuticals.
January 2020, Biotechnology advances,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Improved dual luciferase reporter (DLR) assay to determine the protein stability.
January 2021, Analytical biochemistry,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
A dual-light reporter system to determine the efficiency of protein-protein interactions in mammalian cells.
April 2005, Nucleic acids research,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Assays to determine DNA repair ability.
January 2011, Journal of toxicology and environmental health. Part A,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Dual luciferase gene reporter assays to study miRNA function.
January 2015, Methods in molecular biology (Clifton, N.J.),
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Luciferase Reporter Assays to Determine YAP/TAZ Activity in Mammalian Cells.
January 2019, Methods in molecular biology (Clifton, N.J.),
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Analysis of Regulatory DNA Sequences by Dual-Luciferase Reporter Assays in Ewing Sarcoma.
January 2021, Methods in molecular biology (Clifton, N.J.),
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Improved dual-luciferase reporter assays for nuclear receptors.
January 2010, Current chemical genomics,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Development of luciferase reporter-based cell assays.
June 2006, Assay and drug development technologies,
Huayu Sun, and Sining Wang, and Kebin Yang, and Chenglei Zhu, and Yan Liu, and Zhimin Gao
Standardization of Fluorescent Reporter Assays in Synthetic Biology across the Visible Light Spectrum.
December 2023, ACS synthetic biology,
Copied contents to your clipboard!