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.
Hopkins Marine Station, Stanford University, Pacific Grove, California 93950, USA.
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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