No transgenic rainbow trout produced with sperm incubated with linear DNA.
We attempted to produce transgenic rainbow trout embryos by fertilizing eggs with sperm incubated with linearized plasmids. One experiment was conducted with the construct pBGH7 in the medium MMSF, with or without DMSO, at 2 concentrations of sperm cells and a relatively low concentration of DNA. The DNA was also in contact with the eggs during insemination and during the first minutes of egg activation. The second experiment was conducted with the construct CMVCAT in the medium MMSF, at 2 concentrations of sperm cells and a much higher concentration of DNA. The DNA was also present during the insemination. DNA analyses and dosages of CAT activity did not permit detection of any transgenic fry. However, one result suggests that sperm cells can capture part of the linear DNA in teh conditions tested.
Chourrout
D
D
Laboratory of Fish Genetics, Institut National de la Recherche Agronomique, France.
Perrot
E
E
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000502:physiology*; D005260:Female; D005306:Fertilization; D015870:Gene Expression; D008297:Male; D010957:Plasmids; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology*; D014162:Transfection / Q000379:methods*; D014337:Trout / Q000235:genetics*
D004247:DNA