Expression characterization and promoter activity analysis of the tilapia (Oreochromis niloticus) myosin light chain 3 promoter in skeletal muscle of fish.

A tilapia (Oreochromis niloticus) myosin light chain 3 (Mlc3) promoter region (~4.3 kb) was isolated and characterized. Sequence analysis of the clone revealed high similarity with a tilapia gene encoding the Mlc3 promoter region, exon 1, and intron 1. The clone contained several putative binding sequences for transcription factors, including MEF-2, MYOG, MyoD, PKNOX1, and AREB6. Deletion of a region of the tilapia Mlc3 promoter (801 to -3,881 bp) enhanced promoter activity, as determined by direct injection of a luciferase reporter construct into skeletal muscle of Archocentrus nigrofasciatus. These findings suggest that the region between -801 and -3,881 bp may contain negative regulatory elements. Stable germline transgenic strains of the ornamental fish species A. nigrofasciatus var. carrying the Taiwan coral red fluorescent protein (TcRFP) driven by the Mlc3 promoter were established. F1 adult transgenic A. nigrofasciatus var. exhibited brilliant pink fluorescence in skeletal muscles in the daylight. Therefore, our current study demonstrates the feasibility of using the tilapia Mlc3 promoter to drive fluorescence in new fish species, such as Perciformes.