| 影响水牛胎儿成纤维细胞转染效率的因素 |
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| 引用本文： | 崔奎青,刘庆友,汤刚彬,谢体三,石德顺.影响水牛胎儿成纤维细胞转染效率的因素[J].中国畜牧杂志,2008,44(19). |
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| 作者姓名： | 崔奎青 刘庆友 汤刚彬 谢体三 石德顺 |
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| 作者单位： | 广西大学动物科技学院,广西南宁,530005 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，广西大学校科研和教改项目 |
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| 摘 要： | 实验对电穿孔法和阳离子脂质体法转染水牛胎儿成纤维细胞的条件进行了系统摸索,获得了较好的DNA转染参数。电穿孔法转染时脉冲时间为15ms,电场强度为40～50V/m时,细胞的死亡率处于20%～50%之间,比较适合外源DNA转入水牛胎儿成纤维细胞。将脂质体与DNA的比例控制在8μL:2μg,可获得较高的转染率。相同培养条件下,脂质体法比电穿孔能获得更多的转基因抗性细胞集落,转染DNA的纯度和细胞的状态也明显影响脂质体法的转染效率。
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| 关 键 词： | 转基因 细胞转染 电穿孔法 脂质体法 |
The Factors Affecting Transfection Efficiency of Buffalo Fibroblasts |
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| CUI Kui-qing,LIU Qing-you,TANG Gang-bin,XIE Ti-san,SHI De-shun.The Factors Affecting Transfection Efficiency of Buffalo Fibroblasts[J].Chinese Journal of Animal Science,2008,44(19). |
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| Authors: | CUI Kui-qing LIU Qing-you TANG Gang-bin XIE Ti-san SHI De-shun |
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| Abstract: | The transfection efficiency of electroporation and lipofectin mediated methods was compared. The efficiency of electroporation was line up with the electrical field intensity in which 20%~50% of cells died when the field intensity was set at 40 to 50 V/mm. Employment of 8 μL lipofectin and 2 μg DNA resulted in more cells transfected. Quality of DNA and state of cell were associated with transfection efficiency of lipofectin. At the same culture condition, lipofectin reagent method can obtain more transgenic colonies. |
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| Keywords: | transgenic transfection electroporation lipofectin |
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