Using an improved Tol2 transposon system to produce transgenic zebrafish with epinecidin-1 which enhanced resistance to bacterial infection

Transgenic expression of tilapia hepcidin 1-5 and shrimp chelonianin in zebrafish and their resistance to bacterial pathogens

“…1a) and pTLR-Mylz-EGFP-Mylz-chelonianin (Fig. 1b) were mixed with transposase messenger (m)RNA and then injected into the one-cell stage of zebrafish eggs following our previous report [20]. After microinjection, the eggs were placed in a 28 C incubator, and after 72 h, the presence of green fluorescent color expression was examined by fluorescence microscopy (IX71; Olympus, Tokyo, Japan).…”

“…After microinjection, the eggs were placed in a 28 C incubator, and after 72 h, the presence of green fluorescent color expression was examined by fluorescence microscopy (IX71; Olympus, Tokyo, Japan). For further experiments, we chose one transgenic zebrafish to mate with a wild-type (WT) zebrafish, and mating, breeding, and propagation procedures to the F2 generation followed our previous report [20].…”

“…When the transgenic zebrafish had reached 2 months of age in the F4 generation, they were formalin-fixed, and 4-mm paraffinembedded sections were prepared. Immunohistochemistry was performed as described previously [20].…”

“…TH1-5 and chelonianin might be effective in controlling bacterial pathogens in fish by utilizing transgenic fish technology. To further test this hypothesis, we separately transferred the coding region of TH1-5 and shrimp chelonianin under the control of the zebrafish muscle-specific mylz2 promoter [20] into zebrafish (D. rerio) and studied the resistance of the resulting transgenic zebrafish after an injection of Vibrio vulnificus (204) (a gram-negative bacterium) and Streptococcus agalactiae (SA48) (a gram-positive bacterium). In this article, we demonstrate that F2 transgenic zebrafish expressing TH1-5 or shrimp chelonianin exhibited resistance to infection by V. vulnificus (204) and S. agalactiae (SA48).…”

Transposable Elements in Fish Chromosomes: A Study in the Marine Cobia Species

“…In fact, the physical mapping of TEs, indispensable for its characterization, are available so far for only 32 fish species [Ferreira et al, 2011b], preventing a broader understanding of their distribution patterns and evolutionary role in phylogenetic analyses. Furthermore, long TEs as Tol1 and Tol2, both belonging to the hAT family of transposons, have provided a potential and powerful tool for genetic studies in transgenesis, mutagenesis, gene trapping, and enhancer trapping [Miskey et al, 2005;Parinov and Emelyanov, 2007;Peng et al, 2010].…”