| 延边黄牛体细胞克隆融合条件的研究 |
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| 引用本文： | 杨月春,李钟淑,王士勇,方南洙.延边黄牛体细胞克隆融合条件的研究[J].延边大学农学学报,2009,31(3):153-158. |
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| 作者姓名： | 杨月春 李钟淑 王士勇 方南洙 |
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| 作者单位： | 延边大学农学院,吉林,龙井,133400 |
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| 基金项目： | 延大研科合字，国家自然科学基金项目 |
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| 摘 要： | 以体外成熟培养22-24h的延边黄牛卵母细胞作为受体，颗粒细胞为供核细胞，挤压法去核，注入供体颗粒细胞到卵黄周隙中，甘露醇融合液中施加20μs时长的电脉冲刺激使之融合，复合化学方法激活，从融合前恢复培养时间、融合电场强度、甘露醇浓度等3个方面研究延边黄牛体细胞克隆的适宜融合条件．结果表明，2．5h处理组的融合率（70．9％）显著高于4．0h处理组（58．2％，P〈0．05），2．5h处理组重组胚的卵裂率（77．9％）显著高于其它4组（P〈O．05），其它各组间差异不显著（P〉0．05）；融合电场强度1100V／cm处理组的融合率（86．4％）显著高于900V／cm处理组（67．1％）和1200V／cm处理组（75．2％，P〈O．05），重组胚的卵裂率（84．6％）显著高于900V／cm处理组（69．6％，P〈O．05），囊胚发育率（26．1％）显著高于900V／cm处理组（10．0％），1200V／cm处理组（8．0％，P〈O．05）；不同浓度的甘露醇对融合率和重组胚的卵裂率没有明显的影响，但是0．28mol／L组囊胚发育率（25．3％）显著高于0．25mol／L组（15．3％，P〈0．05）．因此，适合延边黄牛体细胞克隆的融合条件为融合前恢复培养2．5h，0．28mol／L甘露醇作为融合液，电场强度1100V／cm．
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| 关 键 词： | 延边黄牛 体细胞克隆 电融合 重组胚 |
Study of fusion condition of somatic cell cloning of Yanbian Yellow Cattle |
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| YANG Yue-chun,LI Zhong-shu,Wang Shi-yong,FANG Nan-zhu.Study of fusion condition of somatic cell cloning of Yanbian Yellow Cattle[J].Journal of Agricultural Science Yanbian University,2009,31(3):153-158. |
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| Authors: | YANG Yue-chun LI Zhong-shu Wang Shi-yong FANG Nan-zhu |
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| Institution: | ＊ ( Agriculture College of Yanbian University, Longjing Jilin 133400,China) |
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| Abstract: | Make the oocytes IVM 22--24 h as recipient cells, granulosa cells as donor cells, by acupressure enucleated oocytes method putting donor granulosa cells into yolk space, fu- sion with mannitol solution imposed 20 μs electrical pulse length, activated with chemical activator compound,investigate the suitable fusion conditions of somatic cell cloning of Yan- bian Yellow Cattle from recovery time before fusion, electric field strength in fusion and concentration of rnannitol The results showed that fusion rate of 2.5 h treatment group （70.9%） was significantly higher than 4.0 h treatment group （58.2% ,p〈0.05）, cleavage rate of 2.5 h treatment group （77.9%） was significantly higher than the other 4 treatment groups, there was no significant difference in the 4 treatment groups （p〉0.05）; fusion rate of 1 100 V/cm treatment group （86. 4%） was significantly higher than 900 V/cm treatment group （67.1%） and 1 200 V/cm treatment group （75.2%,p〈0.05）, cleavage rate of 1 100 V/cm treatment group （84. 60//00） was significantly higher than 900 V/cm treatment group （69. 6%, p〈0. 05）, blastocyst rate of ll00V/cm treatment group （26.1%） was significantly higher than 900 V/cm treatment group （10.0%） and 1 200 V/ cm treatment group （8.0 %, p〈0.05） . effects of different concentrations of mannitol on fu- sion rate and cleavage rate were not significantly different, but the blastocyst rate of 0.28 mol/L treatment group （25.3 %） was significantly higher than 0.25 mol/L treatment group （15.3%, p~0. 05）. Concluded that suitable fusion conditions of somatic cell cloning in Yanbian Yellow Cattle was recovery time before fusion in 2.5 h, the concentration of with 0.28 mol/L,electric field strength in fusion with 1 100 V/cm. |
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| Keywords: | Yanbian Yellow Cattle somatic cell cloning electric fusion reconstructed em- bryos |
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