ZFIN ID: ZDB-PUB-010131-23
Inheritance, expression and screening of reporter genes in transgenic zebrafish
Gibbs, P.D.L.
Date: 1992
Source: Ph.D. Thesis : (Thesis)
Registered Authors: Gibbs, Patrick
Keywords: Biological sciences
MeSH Terms: none
PubMed: none
Techniques for the high frequency transfer of foreign DNA into the germ line of zebrafish were established. Plasmid DNAs containing combinations of the reporter genes chloramphenicol acetyltransferase (CAT), B-galactosidase (BGAL), hygromycin phosphotransferase (HPT), neomycin phosphotransferase (NPT), firefly luciferase and Drosophila P element sequences were used to make 18 lines of fish exhibiting stable inheritance. Of 17 lines tested for insertional mutagenesis, all were homozygous viable without apparent phenotypic change through embryogenesis. Of the reporter genes CAT, BGAL, NPT and luciferase, expression of CAT and luciferase could be measured transiently in the founder (G0) generation, while after germ line passage only low levels of luciferase expression could be detected. This low level expression could be increased by application of 5-azacytidine, a methylation inhibitor, thus suggesting that integrated expression is suppressed by DNA methylation. A novel photographic screen was developed which utilized luminescence of embryos expressing the luciferase transgene while raised over film. This screen may be useful for large scale genetic analysis of the zebrafish.
Ph.D. Thesis, Washington State University