Reporter gene assays are important tools for evaluating gene expression. A frequently used assay measures the activity of β-galactosidase (β-gal) expressed from lacZ in plasmid or genomic constructions. Such constructions are often used to interrogate the ability of DNA (query DNA), potentially encoding a transcription factor, to regulate in trans the expression of a promoter fused to the reporter lacZ. Query DNA is frequently inserted into a second plasmid within the α-subunit of β-gal, interrupting its function. However, this plasmid can induce up-expression of β-gal even when void of query DNA, leading to confusion between artifact and authentic regulation.
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http://dx.doi.org/10.1016/j.ab.2011.10.019 | DOI Listing |
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