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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

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【作者】 王亮; 彭涛; 朱海; 吕自力; 帅志强; 刘婷婷; 高红; 蔡涛; 曹旭; 汪汉卿;

【Author】 WANG Liang~(1,3,4),PENG Tao~2,ZHU Hai~2,LV Zi-li~2,SHUAI Zhi-qiang~2,LIU Ting-ting~2,GAO Hong~2,CAI Tao~2,CAO Xu~3 and WANG Han-qing~(1*)(1.Key Laboratory for Nature Medicine of Gansu Province,Lanzhou Institute of Chemical Physics,Chinese Academy of Sciences,Lanzhou 730000,China;2.Xinjiang Goldcattle Bio.Inc.,Urumqi 8300263,China;3.Xinjiang Institute of Physics and Chemistry,Chinese Academy of Sciences,Urumq 830011,China;4.Graduate School of the Chinese Academy of Sciences,Beijing 100039,China)

【机构】 中科院兰州化学与物理研究所甘肃省天然药物重点实验室; 新疆金牛生物科学院; 中科院新疆分院理化技术研究所; 中科院兰州化学与物理研究所甘肃省天然药物重点实验室 兰州730000; 乌鲁木齐830011; 中科院北京研究生院; 北京100039; 乌鲁木齐830026; 兰州730000;

【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.

【基金】 国家自然科学基金项目(30471242);中国科学院百人计划项目资助
  • 【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
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