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A newly isolated Japanese flounder ( Paralichthys olivaceus ) &amp;beta;-actin promoter and its derivative compact construct Po&amp;beta;-actin&amp;Delta;&amp;minus;1080/&amp;minus;801&amp;Delta;&amp;minus;500/&amp;minus;201 have recently been demonstrated to promote ectopic gene expression in cell lines. Different Po&amp;beta;-actin promoter deletion mutants were constructed and functionally characterized. Mutational analyses by dual-luciferase detected that three regulatory elements, including one enhancer (&amp;minus;1399/&amp;minus;1081) and two silencers (&amp;minus;1080/&amp;minus;801, &amp;minus;500/&amp;minus;201) in the first intron. The sequence located at &amp;minus;1399/&amp;minus;1081 was determined to significantly affect promoter activity. Additionally, the first exon (&amp;minus;1489/&amp;minus;1400) could also remarkably promote the &amp;beta;-actin promoter activity. In the following transduction application, we removed the two silencers and generated a compact reconstruct promoter/enhancer (Po&amp;beta;-actin&amp;Delta;&amp;minus;1080/&amp;minus;801&amp;Delta;&amp;minus;500/&amp;minus;201), which exhibited relatively stronger promoter activity compared with Po&amp;beta;-actin. Furthermore, the green fluorescent protein (GFP) transgenic stable flounder cell line was obtained by the reconstructed Po&amp;beta;-actin&amp;Delta;&amp;minus;1080/&amp;minus;801&amp;Delta;&amp;minus;500/&amp;minus;201 promoter. Our study provided the potential application of Japanese flounder &amp;beta;-actin, particularly Po&amp;beta;-actin&amp;Delta;&amp;minus;1080/&amp;minus;801&amp;Delta;&amp;minus;500/&amp;minus;201, in ectopic gene expression in the future.

Functional Analysis of the Promoter Region of Japanese Flounder (Paralichthys olivaceus) β-actin Gene: A Useful Tool for Gene Research in Marine Fish