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Electroporation as an effective means of introducing DNA into abalone (Haliotis rufescens) embryos.

作者信息

Powers D A, Kirby V L, Cole T, Hereford L

机构信息

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

出版信息

Mol Mar Biol Biotechnol. 1995 Dec;4(4):369-75.

PMID:8541986
Abstract

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.

摘要

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