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鸡胚培养及活体电转基因方法的建立 被引量：8

Establishment of chicken embryo culture system and in vivo electroporation methods
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摘要 目的建立和优化鸡胚胎带壳培养(in ovo culture)、去壳培养(ex ovo culture)和活体原位电转基因(in vivo electroporation)等技术。方法鸡胚孵化第2～3天,带壳培养至2～3 d和去壳培养至5～6d,分别在脊髓和大脑视顶部位,在电压18V、电流60mA、间隔90ms和电脉冲6次(60ms/次)的条件下进行定时定位活体电转基因。结果带壳和去壳培养样本数分别为20个和10个,成活率分别是85%和80%,胚胎发育至2～3 d和5～6 d在脊髓和视顶部位转染样本数分别为11个和10个,阳性表达率为54.5%和60%。结论成功建立和优化了鸡胚培养和活体定时定位电转基因的方法。 Objective To establish the methods of in ovo and ex ovo culture of chicken embryo as well as in vivo eleetroporation. Methods Fertile eggs were incubated for two to three days （E2-E3）, and in ovo electroporation in spinal cord at E2-E3, and ex ovo clectroporation in the rectum of brain at E5-E6 for spatial and temporal gene transformation with the parameter such as volt 18V, current 60mA, pause 90ms, and pulse 60ms for six times were carried out. Results The number of samples were 20 eggs for in ovo culture and 10 eggs for ex ovo culture. The survival rate of the embryos at E2-E3 was 85% for in ovo culture and 80% for ex ovo culture. The number of samples were 11 in spinal cord at E2-E3 （in ovo electroporation） and 10 in brain tectum at ES-E6 （ex ovo electroporation）. The positive electroporation rate was 54.5% in spinal cord at E2-E3 （in ovo） and 60% in brain tectum at E5-E6 （ex ovo）. Conclusion The methods of in ovo and ex ovo culture of chicken embryos and in vivo electroporation are established successfully.
作者 杨慈清 毛会丽 林俊堂
机构地区 新乡医学院生命科学技术系
出处 《解剖学报》 CAS CSCD 北大核心 2012年第4期564-568,共5页 Acta Anatomica Sinica
基金 国家自然科学基金资助项目(31000475)
关键词 发育 培养 活体电转 鸡胚 Development Culture In vivo electroporation Chicken embryo
分类号 Q954.48 [生物学—动物学]
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参考文献13

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共引文献2

同被引文献59

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解剖学报
2012年 第4期

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