Transgenic Zebrafish Expressing Chicken Lysozyme Show Resistance against Bacterial Diseases |
| |
Authors: | Ryosuke Yazawa Ikuo Hirono Takashi Aoki |
| |
Institution: | (1) Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, 108-8477 Minato, Tokyo, Japan |
| |
Abstract: | We established a transgenic zebrafish strain expressing chicken lysozyme gene under the control of the Japanese flounder keratin
gene promoter, and investigated its resistance to a pathogenic bacterial infection. To generate the lysozyme transgenic construct,
Japanese flounder keratin promoter was linked to both the hen egg white (HEW) lyoszyme gene and green fluorescence protein
(GFP) gene used as a selection marker for the transgenic strains, in a recombinant plasmid. The recombinant plasmid was microinjected
into fertilized zebrafish eggs. In F2 transgenic zebrafish, GFP expression was strong in the epithelial tissues, liver and
gill from the embryonic stage to the adult stage. The expressions of HEW lysozyme and GFP mRNA were confirmed in the liver
and skin by RT-PCR. Western blot analysis showed that both HEW lysozyme and GFP were present in protein extracts from the
liver of transgenic zebrafish, but not in protein extracts from the muscle. The lytic activity of protein extracts from the
liver (assessed by a lysoplate assay using Micrococcus lysodeikticus as a substrate) was 1.75 times higher in F2 transgenic zebrafish than in the wild type. In a challenge experiment, 65% of
the F2 transgenic fish survived an infection of Flavobacterium columnare and 60% survived an infection of Edwardsiella tarda, whereas 100% of the control fish were killed by both pathogens. However, the survival rates of the transgenic fish were
not significantly higher when higher concentrations of bacteria were used. |
| |
Keywords: | disease resistance keratin promoter lysozyme transgenic fish zebrafish |
本文献已被 PubMed SpringerLink 等数据库收录! |
|