( ...)Genetic Reporter Systems

Genetic reporter systems have developed into an essential tool for examining regulatory promoter and enhancer sequences as well as transcription factors. In most cases, the element under investigation (promoter, enhancer) is cloned together with the reporter gene in an expression vector, which is subsequently used to transfect cells. Quantification of the reporter indirectly provides information on the transcription activity of the element under investigation. Quantification can take place by detecting the corresponding RNA, the reporter protein, or by measuring the enzyme activity of the reporter protein. When the reporter system is selected, care must be taken to ensure that the reporter gene does not influence the physiology of the transfected cells and that the gene is not already endogenously expressed in the examined cells.

Reporter systems are also often used as a standard to compare the transfection efficiency of different transfection experiments. In this case, the control reporter system usually contains a constitutive promoter and a reporter gene, which is different to the reporter gene used by the element under examination. Such a reporter system can also be used to optimize electroporation parameters.

Chloramphenicol Acetyltransferase (CAT)

Chloramphenicol Acetyltransferase (CAT) This enzyme comes from microorganisms and catalyzes the transfer of acetyl groups from acetyl coenzyme A to chloramphenicol. With the CAT assay, the CAT-containing lysates of transfected cells are incubated with ¹⁴C-chloramphenicol, which is then acetylated. Acetylated and non-acetylated ¹⁴C-chloramphenicol is then separate
'"/>

Source:

Page: All 1 2 3 4
Related biology technology :

1. Map and Link Human Genetic Disorders with SSLP Analysis
2. Genetic Reporter Systems
3. Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing
4. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
5. The iCycler iQ and Amplifluor Detection Systems for Analysis of Genetically Modified Organisms in Foodstuffs, Rev A
6. Analysis of Bacterial Assemblage Genetic Diversity in Environmental Samples Using the DCode System
7. Monitoring Bacterial Genetic Diversity in a Freshwater Lake Using TTGE and DNA Sequence Analysis
8. Rapid, High Throughput Mutation Discovery Using Fluorescent SSCP on the ABI PRISM 3100 and 3100Avant Genetic Analyzers
9. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
10. Stable HeLa Luciferase Reporter Cell Lines Expressing GAL4 Fusion Transactivators
11. Versatile Reporter Vectors for Monitoring Viral Transduction