Gene Analyses--Sunny Side Up!

Chemiluminescent Gene Reporter Systems Features of Luminometers How is the expression of a given gene controlled? That is a question asked of many in the biological and medical sciences. "Reporter genes," which encode quantifiable enzymes or proteins, are often employed by researchers seeking to better understand how the expression of genes of interest is controlled. In these assays, the coding region of the gene under study is replaced with the sequence that encodes the reporter gene. Report

Written byDeborah Wilkinson
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A reporter gene must fulfill several basic criteria in order to be useful. A simple, cost-effective assay for quantifying the reporter protein or activity should be available, and the amount of reporter gene protein must correspond to the amount of reporter gene mRNA transcribed. Endogenous protein or enzyme activity must be absent or minimal in target cells. The assay should also have a broad linear range to enable analyses of both large and small changes in gene expression. Further, expression of the reporter should not alter the physiology of the recipient cells. An additional concern is the relative sensitivity of the assay. Sensitivity becomes an issue when attempting to analyze low levels of gene expression, especially in cell lines that transfect poorly. The sensitivity of each reporter gene assay does not only depend on the sensitivity of the detection method itself. It is also a function of numerous cell-line-specific factors, ...

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