Replication, expression, and fate of foreign DNA during embryonic and larval development of the African catfish (Clarias gariepinus).
The transfer of exogenous DNA in fish represents a powerful strategy to study the regulation of gene expression in vivo. The African catfish (Clarias gariepinus) was chosen for this study because of its scientific and economic importance due to its easy husbandry, its short developmental period, and its value as a protein source in Africa and Asia. Fertilized eggs (1- and 2-cell stage) were cytoplasmatically injected with either supercoiled or linearized plasmids harboring the fusion genes encoding beta-galactosidase (lacZ) or luciferase (Luc) without a promoter or fused to the promoter/enhancer of human cytomegalovirus (CMV). Replication of the exogenous DNA peaked at 4 hours (early gastrula) and again at 2 days (which corresponds to the developmental stage of yolksac resorption). Foreign DNA persisted during embryogenesis, and it was still detectable 8 months after injection. In vivo transient expression of both CMV fusion genes was mosaic and peaked within 24 hours after DNA injection. Transient expression of the luciferase reporter gene could be detected with a much higher sensitivity than the lacZ gene. These data establish African catfish as a suitable in vivo assay system, and they confirm the luciferase reporter gene as a high quality reporter gene in fish.
Volckaert
F A
FA
Laboratory of Ecology and Aquaculture, Katholieke Universiteit Leuven, Belgium.
Hellemans
B A
BA
Galbusera
P
P
Ollevier
F
F
Sekkali
B
B
Belayew
A
A
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D015139:Blotting, Southern; D002397:Catfishes / Q000196:embryology / Q000235:genetics*; D046508:Culture Techniques; D017931:DNA Primers; D004261:DNA Replication; D004274:DNA, Recombinant / Q000235:genetics*; D005260:Female; D018014:Gene Transfer Techniques; D007814:Larva / Q000378:metabolism; D008297:Male; D008845:Microinjections; D008969:Molecular Sequence Data
D017931:DNA Primers; D004274:DNA, Recombinant