Detection of Recombinant Protein Based on Reporter Enzyme Activity: Chloramphenicol Acetyltransferase

Genetic engineering technologies allow the construction of genetic chimeras between the promoter region of a gene of interest and a reporter gene as a means to study the regulation of eukaryotic gene expression at the transcriptional level. The genetic constructs in plasmid form are delivered back into cells to measure the expression of the reporter gene. A good reporter gene product possesses the following characteristics: the enzymatic activity is heat stable and protease resistant, and corresponds well to the strength of the promoter; background and/or interfering enzymatic activities are not present in mammalian cells; and, simple, sensitive, reproducible, and convenient enzymatic or immunological assays of the reporter gene product are available for the assessment of the promoter activity ( 1 ). For many applications, the bacterial chloramphenicol acetyltransferase (CAT) satisfies these criteria ( 2 ). These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.