Electroporation as an effective means of introducing DNA into abalone (Haliotis rufescens) embryos.
Recombinant plasmid containing the Drosophila beta-actin promoter coupled to a beta-galactosidase cassette was linearized and introduced in fertilized eggs of the red abalone (Haliotis rufescens) by electroporation. Fertilized abalone eggs tolerated electroporation well with larval survival rates between 70% and 84% of that for non-electroporated siblings. Dot blot and Southern blot analysis were used to detect if abalone retained the foreign gene at various developmental stages. The inserted construct was retained in 70% to 100% of all abalone sampled with an average of 72% retention in the three- to seven-month-old juveniles. Maximal DNA uptake and retention was observed in abalone electroporated at 30-40 min after fertilization. Southern hybridization analysis suggested that the inserted vector was in head-to-tail concantermers integrated in the abalone genome. This preliminary study demonstrates that electroporation is an efficient means of transferring foreign DNA into abalone embryos.
Powers
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Hopkins Marine Station, Stanford University, Pacific Grove, California 93950, USA.
Kirby
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Cole
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Hereford
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D000199:Actins / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D004247:DNA / Q000378:metabolism; D018274:Electroporation; D018014:Gene Transfer Techniques; D017344:Genes, Insect; D008974:Mollusca / Q000196:embryology / Q000235:genetics*; D010957:Plasmids; D015183:Restriction Mapping; D001616:beta-Galactosidase / Q000235:genetics
D000199:Actins; D004247:DNA; D001616:beta-Galactosidase