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转染方法对肌肉生长抑制素基因敲除载体转染效率的影响 被引量：1

Influence of transfecting methods on the efficiency of myostatin knockout vector
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摘要 本研究主要探讨转染方法对不同细胞系的转染效果。实验采用电穿孔和脂质体转染方法,将线性化的绵羊肌肉生长抑制素基因(myostatin)药物正负筛选(PNS)打靶载体pLoxp-1.4K-4.3K DNA导入体外培养的绵羊胎儿和成年绵羊耳部成纤维细胞中,而后采用G418和GANC进行药物抗性细胞克隆筛选和培养。结果发现,脂质体法对myostatin基因敲除载体的转染效果明显优于电击法,但电穿孔法对细胞类型的依赖性较小;采用与载体基因型相同的细胞系,两种转染方法的转染效率均略高于对照成年绵羊成纤维细胞系。 The transfecting efficiency of electroporation and lipofectamin on different fibroblasts cultured in vitro was compared in this study. Sheep fetal and adult fibroblasts were transfected with sheep myostatin replacement vector pLoxp-1.4K-4.3K by either electroporation or lipofectamin, then cultured in G418＋/GANC＋ drug selection medium. Calculated from the number of G418＋/GANC＋ resistant cell clone, the efficiency of lipofectamin transfection method was higher than that of the electroporation, however, the transfecting efficiency of electroporation was independent of cell type. The efficiency of DF1 cell lines （the genome identical with the constructed vector） was higher than that of the DF2 （the common fibroblasts） in the two methods.
作者 李湘萍 黄时海 石德顺 李宁
机构地区 广西大学动物繁殖研究所 广西大学生命科学与技术学院 中国农业大学农业生物技术国家重点实验室
出处 《广西农业生物科学》 CAS CSCD 2007年第B06期27-31,共5页 Journal of Guangxi Agricultural and Biological Science
基金 广西青年科学基金项目(桂科青0339006) 国家自然科学基金项目(30660126)
关键词 脂质体法 电穿孔法 肌肉生长抑制素基因 基因敲除 lipofectamin transfection electroporation myostatin gene knockout
分类号 Q78 [生物学—分子生物学]
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参考文献10

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同被引文献14

  • 1杨素芳,石德顺,韩杰,韦英明,韦精卫,陆凤花,牛向丽.供体核种类对水牛核移植效果的影响[J].广西农业生物科学,2004,23(3):233-237. 被引量：16
  • 2王海嵘,顾春红,钟济华,王婷,韩洁英,陈芳源,欧阳仁荣.流式细胞术结合G418筛选重组质粒稳定转染细胞[J].诊断学理论与实践,2006,5(1):52-55. 被引量：9
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广西农业生物科学
2007年 第B06期

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