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High efficiency gene transfer in rainbow trout (Salmo gairdneri Rich.) by microinjection into egg cytoplasm |
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| Affiliation: | 1. University of Bristol, School of Veterinary Sciences, Regenerative Medicine Laboratory, Biomedical Science Building, University Walk, Bristol BS8 1TD, United Kingdom;2. University of Bristol, School of Clinical Sciences, Regenerative Medicine Laboratory, Biomedical Science Building, University Walk, Bristol BS8 1TD, United Kingdom;3. University of Cambridge, Department of Clinical Neurosciences, Cambridge Centre for Brain Repair, E.D. Adrian Building, Forvie Site, Robinson Way, Cambridge CB2 0PY, United Kingdom;4. University of Cambridge, Department of Physiology Development and Neuroscience, Anatomy Building, Downing St, Cambridge CB2 3DY, United Kingdom;5. University of Bristol, School of Veterinary Sciences, Langford House, Langford, North Somerset BS40 5DU, United Kingdom |
| Abstract: | A plasmid containing human growth hormone cDNA sequence was injected into cytoplasm of fertilized rainbow trout (Salmo gairdneri Rich.) eggs before first cleavage. Survival rates at hatching (stage of analysis) were high (average: 77%; range: 50–100%). Foreign DNA was detected in pooled and individual embryos either by dot blot or by southern electrophoresis procedures using the labelled plasmid as probe; it comigrated with the high molecular weight DNA and was therefore most likely integrated in the trout genome. Transformation yields were higher with linearized than with circular plasmid (75% versus 40%). These data suggest that transgenic trout can be routinely obtained in large proportions. |
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