LipofectamineTM和Fugene-6介导GFP基因转染绵羊胎儿成纤维细胞的比较研究

›› 2008, Vol. 10 ›› Issue (1): 108-112.

LipofectamineTM和Fugene-6介导GFP基因转染绵羊胎儿成纤维细胞的比较研究

马玉珍[1,2] 王瑞[2] 闫真[2] 王利民[2] 刘东军[2] 夏国良[1]

[1]中国农业大学生物学院,北京100094 [2]内蒙古大学哺乳动物繁殖生物学与生物技术教育部重点实验室,呼和浩特010021

收稿日期:2007-12-29 修回日期:2008-01-11 出版日期:2008-02-15 发布日期:2009-07-30
作者简介:马玉珍|副研究员|博士|主要从事动物生殖生物学和生物技术研究。Tel：0471-4992443；E-mail：mayz@imnu．edu．cn
基金资助:
国家自然科学基金项目（30060059）和新疆建设兵团博士基金项目（05JC03）资助.

Comparison Between Sheep Fetal Fibroblast Cells transfected with |GFP by Lipofectamine^TM and by Fugene-6

MA Yu-zhen, WANG Rui,YAN Zhen, WANG Li-min| LIU Dong-jun| XIA Guo-liang

1. College of Biological Science, China Agricultural University, Beijing 100094|2. Key Laboratory of Mammal Reproduetive |Biology and Biotechnology, Ministry of Education, Inner Mongolia University, Huhhot 010021, China

Received:2007-12-29 Revised:2008-01-11 Online:2008-02-15 Published:2009-07-30

摘要/Abstract

摘要：

分别用Lipofectamine^TM和Fegene-6介导质粒pEGFP-C1转染体外培养的绵羊胎儿成纤维细胞（sheepfetal fibroblast cells,sFFCs）,比较了DNA浓度、转染试剂用量以及细胞暴露于DNA、转染试剂中作用时间对转染效率的影响,通过含G418的DMEM/F12培养液筛选得到转基因单克隆细胞。结果表明脂质体转染试剂Lipofectamine^TM转染效率优于Fegene-6。另外,对转基因细胞进行了染色体核型分析。结果表明,转基因细胞中二倍体核型占74.5%,与对照组比较没有显著性差异。以上研究为其他基因转染sFFCs以及利用体细胞克隆法生产转基因绵羊提供了参考依据。

关键词: 绵羊胎儿成纤维细胞脂质体转基因染色体

Abstract:

Sheep fetal fibrobast cell were transfected with pEGFP-C1 by using Lipofectamine^TM or Fugene-6 substance respectively. The impacts of DNA concentration, concentration of transfeetion agents, exposure time and incubation time on transfection efficiency were carefully compared. The transfected cells were incubated in DMEM/F12 media containing G418. The results showed that the efficiency of transgenic with LipofectamineTM were excelled Fugene-6. Chromosome analysis shows no remarkable difference between transgenic and normal cells. LipofectamineTM transfection method developed in this paper can be easily adopted by most research labs and will provide a basic reference for transfection of sheep fetal flbroblast ceils with other gene and transgenie nuclear transfer.

Key words: sheep fetal fibroblast cell , liposome , gene transfer, chromosome

中图分类号:

Q813 Q959.842

马玉珍,王瑞,闫真,王利民,刘东军,夏国良. LipofectamineTM和Fugene-6介导GFP基因转染绵羊胎儿成纤维细胞的比较研究[J]. , 2008, 10(1): 108-112.

MA Yu-zhen, WANG Rui,YAN Zhen, WANG Li-min| LIU Dong-jun| XIA Guo-liang. Comparison Between Sheep Fetal Fibroblast Cells transfected with |GFP by Lipofectamine^TM and by Fugene-6[J]. , 2008, 10(1): 108-112.