Illustration from Scientific Research

To functionally test the necessity of a predicted RFX-binding site for gene activation, the 5′ UTR and proximal promoter of the spag6 gene from S. rosetta was cloned in front of an anoLuc open reading frame, which was codon-optimized for S. rosetta. A second reporter construct was made in which the predicted RFX-binding site (marked in red) was mutated (italicized letters). As an internal normalization step, the plasmid also encodes the Firefly luciferase under strong expression from the S. rosetta actin promoter.

DOI