We examined the efficiency of electroporation for the delivery of plasmid into the skeletal muscle and also examined the subsequent secretion of recombinant protein into the circulation system, using zebrafish, Japanese flounder, and bubble-eye goldfish. The expression area of GFP fluorescence was markedly expanded by electroporation. Introduced plasmid was retained in the muscle cells and continued to express GFP for at least 180 days in zebrafish, indicating the long lifespan of plasmid DNA in the muscle cell. Luciferase and a fusion of growth hormone (GH) and luciferase were secreted into the blood from muscle electroporated with CMV:secNluc and CMV:GH-Luc plasmids, respectively, indicating that recombinant proteins such as peptide hormones can be supplied to the blood by plasmid electroporation into muscle. Interestingly, luciferase activity was detected from fertilized eggs laid by zebrafish females that had been electroporated with CMV:secNluc, indicating that maturing oocytes incorporated recombinant protein from the blood stream that had been secreted from the muscle. The plasmid electroporation system reported here also may work for the delivery of recombinant proteins, such as Cas9, into the oocytes. Since the GH-Luc fusion protein was detected in the lymph of the eye-sac of bubble-eye goldfish, this fish may be useful for the production of recombinant protein.

中文翻译：

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通过电穿孔将质粒递送到鱼骨骼肌中以重组蛋白分泌和卵母细胞摄取。

我们使用斑马鱼，日本比目鱼和泡眼金鱼，研究了电穿孔将质粒递送至骨骼肌的效率，还研究了重组蛋白随后向循环系统中的分泌。通过电穿孔，GFP荧光的表达区域显着扩大。引入的质粒保留在肌肉细胞中，并在斑马鱼中持续表达GFP至少180天，这表明质粒DNA在肌肉细胞中的寿命很长。萤光素酶以及生长激素（GH）和萤光素酶的融合物分别通过CMV：secNluc和CMV：GH-Luc质粒电穿孔的肌肉分泌到血液中，表明重组蛋白（如肽激素）可以通过质粒提供给血液电穿孔进入肌肉。有趣的是 从用CMV：secNluc电穿孔的斑马鱼雌性产下的受精卵中检测到萤光素酶活性，表明成熟的卵母细胞掺入了从肌肉分泌的血流中获得的重组蛋白。此处报道的质粒电穿孔系统也可用于将重组蛋白（例如Cas9）输送到卵母细胞中。由于在泡泡眼金鱼的眼囊淋巴中检测到了GH-Luc融合蛋白，因此该鱼可用于生产重组蛋白。此处报道的质粒电穿孔系统也可用于将重组蛋白（例如Cas9）输送到卵母细胞中。由于在泡泡眼金鱼的眼囊淋巴中检测到了GH-Luc融合蛋白，因此该鱼可用于生产重组蛋白。此处报道的质粒电穿孔系统也可用于将重组蛋白（例如Cas9）输送到卵母细胞中。由于在泡泡眼金鱼的眼囊淋巴中检测到了GH-Luc融合蛋白，因此该鱼可用于生产重组蛋白。