›› 2012, Vol. 43 ›› Issue (4): 564-568.doi: 10.3969/j.issn.0529-1356. 2012.04.023

• 技术方法 • 上一篇    下一篇

鸡胚培养及活体电转基因方法的建立

杨慈清; 毛会丽; 林俊堂*   

  1. 新乡医学院生命科学技术系,河南省医用组织再生重点实验室,河南 新乡 453003
  • 收稿日期:2011-05-02 修回日期:2011-07-10 出版日期:2012-08-06
  • 通讯作者: 林俊堂

Establishment of chicken embryo culture system and EM>in vivo/EM> electroporation methods

  1. Department of Life Science and Technology, Key Laboratory of He’nan Province for Medical Tissue Regeneration. Xinxiang Medical University, He’nan Xinxiang 453003,China
  • Received:2011-05-02 Revised:2011-07-10 Online:2012-08-06
  • Contact: LIN Jun-tang

关键词: 发育, 培养, 活体电转, 鸡胚

Abstract: Objective To establish the methods of EM>in ovo/EM> and EM>ex ovo/EM> culture of chicken embryo as well as EM>in vivo/EM> electroporation. 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 electroporation in the tectum 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 EM>in ovo/EM> culture and 10 eggs for EM>ex ovo/EM> culture. The survival rate of the embryos at E2-E3 was 85% for EM>in ovo/EM> culture and 80% for EM>ex ovo/EM> culture. The number of samples were 11 in spinal cord at E2-E3 ( in ovo electroporation) and 10 in brain tectum at E5-E6 ( ex ovo electroporation). The positive electroporation rate was 54.5% in spinal cord at E2-E3 (EM>in ovo/EM> ) and 60% in brain tectum at E5-E6 (EM>ex ovo/EM>). Conclusion The methods of in ovo and ex ovo culture of chicken embryos and in

Key words: Development, Culture, EM>In vivo/EM> electroporation, Chicken embryo

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