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电融合条件对新疆盘羊(Ovis ammon)和转基因(GFP)萨能奶山羊核移植胚胎发育的影响
Influence of Electricfusion Conditions on the Development of Nuclear Transfer Embryos of Xinjiang Wild Ovis ammon and Transgenic (GFP) Saanen Dairy Goat
【摘要】 以新疆野生盘羊(Ovis ammon)和萨能奶山羊转基因(GFP)成纤维细胞为核供体细胞,分别以绵羊卵母细胞和普通山羊卵母细胞为受体细胞进行了核移植,对不同种类核移植重构卵进行电融合处理,发现盘羊重构卵电融合参数为1.4 kV/cm、20μs/次、间隔1 s时的重构卵融合率最高;对于转基因(GFP)萨能奶山羊重构卵的处理,电融合前先放入融合液中于38.5℃下平衡5 min,融合时电融合参数为1.5 kV/cm、20μs/次、间隔1 s时的融合率最高。实验中发现将融合后的两种重构胚胎先置入IVM培养液中于38.5℃下平衡4 h,可降低重构胚胎的死亡率,提高发育质量和卵裂率。
【Abstract】 In this study using the fibroblasts of Xinjiang wild Ovis ammon and the transgenic(GFP) fibroblasts of Saanen dairy goat as donor cells and using the Abattoir-derived oocytes of domesticated sheep and goat as recipient cells the nuclear transfer experiments had been prcessed.During the electricfusion experiments of different kinds of nuclear transfer embryos the high fusion rate of Ovis ammon cell-cytoplasts was found when the electricfusion parameter was 1.4 kV/cm for 20 μsec.For the transgenic cell-cytoplasts of Saanen dairy goat,before electricfusion the cell-cytoplasts were first put into fusionsolution to be balanced for 5 min at 38.5℃,when the electricfusion parameter was 1.5(kV/cm) for 20 μsec,the high fusion rate could be obtained.After electricfusion these two kinds of reconstruction embryos were balanced at 38.5℃ in IVM medium for 4 hours can decrease the death rate of reconstruction embryos and increase the development quality and the cleavage rate of reconstruction embryos.
【Key words】 Transgenic; Nuclear transfer; Electrofusion; Reconstruction embryo; Somatic cloning;
- 【文献出处】 西北农业学报 ,Acta Agriculturae Boreali-Occidentalis Sinica , 编辑部邮箱 ,2006年02期
- 【分类号】S814.8
- 【被引频次】8
- 【下载频次】160