| 英文摘要 |
In this study, we choose a medium-sized freshwater America Cichlidae fish Cichlasoma nigrofasciatum pink mutant, common name as white lion cichlid or pink convict cichlid as trasngenesis target to develop as transgenic fluorescent fish. Due to its good biological characteristics, fecundity and transparent muscle, the transgenic pink convict cichlid with green or red fluorescent muscle will be established by microinjection, electroporation in combined with Tol2 trasnsposon-mediated trasngenesis. The outcome Fluorescent Red Lion Cichlid and Fluorescent Green Lion Cichlid will create new market of fluorescent ornamental fish.
To establish transgenic pink convict cichlid with strong fluorescent muscle which is even visible under the daylight lamp, we choose to use zebrafish strong promoters with muscle specificity and ubiquity, respectively. We had cloned the 2kb muscle -specific myosin-light chain 2 (MCL2) promoter, ubiquitous 2.2kb alpha-actin and 2.5kb beta-actin promoters from zebrafish genomic DNA. The Tol2 transgenesis constructs with muscle-specific MLC2 promoter to express green or cyan fluorescent protein were also finished. This Tol2-mediated transgenesis vector with 2k MLC2 promoter is indeed muscle-specific and fluorescence visible in zebrafish founder. We had successfully selected the F1 offspring of transgenic zebrafish lines with green fluorescent muscle and prove our Tol2-mediated muscle-specific transgenesis vector is workable in establishing transgenic fluorescent ornamental fish. In pink convict, we had obtained seven pairs of naturally selected couple fish to get one-cell stage fertilized eggs for transgenesis and still under selection for transgenic lines |
