| 柔嫩艾美耳球虫电穿孔转染条件的优化 |
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| 引用本文： | 赵世云,苏华荔,刘贤勇,索勋.柔嫩艾美耳球虫电穿孔转染条件的优化[J].中国畜牧兽医,2010,37(11):53-56. |
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| 作者姓名： | 赵世云 苏华荔 刘贤勇 索勋 |
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| 作者单位： | （中国农业大学动物医学院，北京 100193） |
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| 摘 要： | 低转染效率一直是利用基因转染技术研究柔嫩艾美耳球虫的主要屏障。为了进一步提高现有的转染效率，本研究分别利用传统的Bio-Rad Gene Pulser电穿孔仪和新型的NucleofectorⅠ核转仪对柔嫩艾美耳球虫子孢子的转染条件进行了优化。通过对外源DNA浓度、电场强度、环境温度及NucleofectorⅠ核转仪的内置程序这些关键参数的探索，最终发现利用Bio-Rad Gene Pulser电穿孔仪设置电压为2000 V，电容25 μF，利用4 mm电击杯进行电转时，转染效率最高，达到3.8×10-4，此时最佳质粒浓度为50 μg/107子孢子。
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| 关 键 词： | 柔嫩艾美耳球虫 基因转染 转染效率 |
Optimization of Electroporation for Transfection of Eimeria tenella |
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| ZHAO Shi-yun,SU Hua-li,LIU Xian-yong,SUO Xun.Optimization of Electroporation for Transfection of Eimeria tenella[J].China Animal Husbandry & Veterinary Medicine,2010,37(11):53-56. |
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| Authors: | ZHAO Shi-yun SU Hua-li LIU Xian-yong SUO Xun |
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| Institution: | (College of Veterinary Medicine, China Agricultural University, Bejing 100193, China) |
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| Abstract: | Low transfection efficiency has been the major hurdle for gene transfection in Eimeria tenella. In order to overcome this difficulty, the electroporation parameters were optimized for traditional electroporation apparatus of Bio-Rad Gene Pulser and newly developed Amaxa’s Nucleofector I device, respectively. By adjusting series of key parameters, which include the concentration of plasmid DNA, the electric field intensity, the ambient temperature and bult-in programs of Nucleofector I, we found the optimal condition for electroporation of E. tenella sporozoites is at a voltage of 2000 V and a capacitance of 25 μF, using 50 μg plasmids/107 sporozoites and cuvettes with a 4 mm gap, which together resulting in a transfection efficiency of 3.8×10-4. |
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| Keywords: | Eimeria tenella gene transfection transfection efficiency |
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