An enhanced yellow fluorescence protein (eYFP) cassette was introduced in frame with the first ATG in exon 1, followed by a neo cassette flanked by two Frt sites. The neo cassette was removed by Flp-mediated recombination, resulting in a continuous reading frame from the initiating codon, through eYFP and the residual FRT to the second amino acid. No yellow fluorescent signal was detected in any tissue. Real-time PCR and Western blot analysis showed decreased expression in ovaries from homozygous females. (J:166778)