Molecular marine biology and biotechnology vol:3 pages:30-34
Transcriptional regulation of gene expression is often studied by fusing the DNA sequences of interest (and their mutants) with a reporter gene. The resulting hybrid gene is introduced into cells grown in vitro and its transient expression is analyzed. The success of this approach depends strongly on choosing an appropriate reporter gene. In preparation of such studies in fish cells and embryos, we systematically compared the following reporter genes: chloramphenicol acetyltransferase (cat gene), beta-galactosidase (lacZ gene), beta-lactamase (ampR gene) and firefly luciferase (Luc gene) in vitro (EPC cells) and in vivo (transiently transgenic zebrafish). The different reporter gene systems are discussed. Our results establish the luciferase reporter gene as an excellent choice for the intended studies. We show that electroporation is an effective way to introduce foreign DNA into fish cells grown in vitro.