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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
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