| 氯化钠密度梯度离心法制备用于显微注射的外源DNA片段 |
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| 引用本文： | 刘立仁,赵华路,张俊武. 氯化钠密度梯度离心法制备用于显微注射的外源DNA片段[J]. 遗传, 2003, 25(5): 587-590 |
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| 作者姓名： | 刘立仁 赵华路 张俊武 |
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| 作者单位： | 中国医学科学院基础医学研究所 中国协和医科大学基础医学院 医学分子生物学国家重点实验室，北京 100005National laboratory of Medical Molecular Biology,Institute of Basic Medical Science,CAMS&PUMC,Beijing 100005,China |
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| 基金项目： | 国家自然基金(39893320)资助 |
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| 摘 要： | DNA显微注射是生产转基因动物最可靠和最常使用的一种方法，外源DNA的纯度对显微注射的成功起着至关重要的作用。本文介绍用氯化钠密度梯度离心的方法制备用于显微注射的外源DNA片段。与传统的琼脂糖凝胶回收的方法相比较，用此方法制备的外源DNA片段对小鼠受精卵进行显微注射后，受卵体母鼠的胚胎存活率,以及子代小鼠的外源基因整合率均有明显的提高。这一方法可为进一步提高转基因动物的成功率,提供方法学上的参考。Abstract:DNA microinjection is the most popular and reliable method of producing transgenic animals.The purity of foreign DNA plays an important role for the success of microinjection.In this study,we introduced the use of sodium chloride step gradients in fractionating foreign DNA fragment for microinjection.The data demonstrated that,compared with the conventional agarose gel extraction method,NaCl purification scheme of toreign DNA could improve the treated embryo survival and foreign DNA intergration rate markedly.
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| 关 键 词： | microinjection 胚胎存活率 外源基因整合率 Key words 显微注射 氯化钠密度梯度离心 |
| 文章编号： | 0253-9772(2003)05-0587-04 |
| 修稿时间： | 2003-02-14 |
Sodium Chloride Step Gradients Centrifuge to Fractionate Foreign DNA Fragment for Microinjection |
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| Li-Ren Liu,Hua-Lu Zhao,Jun-Wu Zhang. Sodium Chloride Step Gradients Centrifuge to Fractionate Foreign DNA Fragment for Microinjection[J]. Hereditas, 2003, 25(5): 587-590 |
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| Authors: | Li-Ren Liu Hua-Lu Zhao Jun-Wu Zhang |
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| Affiliation: | National laboratory of Medical Molecular Biology, Institute of Basic Medical Science, CAMS&PUMC, Beijing 100005, China. |
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| Abstract: | DNA microinjection is the most popular and reliable method of producing transgenic animals. The purity of foreign DNA plays an important role for the success of microinjection. In this study, we introduced the use of sodium chloride step gradients in fractionating foreign DNA fragment for microinjection. The data demonstrated that,compared with the conventional agarose gel extraction method, NaCl purification scheme of toreign DNA could improve the treated embryo survival and foreign DNA intergration rate markedly. |
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| Keywords: | microinjection sodium chloride step gradients centrifuge embryo survival foreign DNA intergration rate |
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