双壳类软体动物鸡变色龙卵的转染。

R Guerra, P Esponda
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引用次数: 0

摘要

对双壳类软体动物鸡变色龙卵进行了体外转染。采用表达绿色荧光蛋白(GFP)的p-GeneGrip基因构建。成功转染需要去除覆盖在卵表面的果冻层,使用裸DNA后,44.2%的配子转染成功。另一方面,阳离子脂质体(Lipofectamine)和中性脂质体(GenePORTER)作为基因载体。使用Lipofectamine后,转染率为35.6%,使用GenePORTER后转染率为41.4%。荧光分析显示外源基因主要存在于卵细胞质中,激光共聚焦显微镜显示外源基因也存在于细胞核中。此外,PCR分析表明,从处理过的卵中提取的DNA中出现了外源DNA。这种简单的转染软体动物卵的方法对未来具有商业价值的物种的生物技术应用很有意义。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Transfection of eggs in the bivalve mollusc Chamelea gallina (Bivalvia, Veneridae).

Eggs from the bivalve mollusc Chamelea gallina were transfected in vitro. The p-GeneGrip gene construction that expresses the green fluorescent protein (GFP) was employed. It was necessary to remove the jelly coat which covers the egg surface for a successful transfection, and then 44.2% of gametes appeared transfected after using naked DNA. On the other hand, cationic liposomes (Lipofectamine) and neutral lipids (GenePORTER) were employed as gene vectors. After the employ of Lipofectamine 35.6% of eggs were transfected and 41.4% after using GenePORTER. Fluorescence analysis showed that the foreign gene appeared principally located in the egg cytoplasm, but laser confocal microscopy showed that it was also present in the nucleus. Furthermore, PCR analysis demonstrated that the foreign DNA appeared in the DNA extracted from the treated eggs. This simple method for the transfection of mollusc eggs would be interesting for future biotechnological applications in species of commercial interest.

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