[1]时晓芳),闫红霞),宋安营),等.稳定转染pEGFP4.1N乳癌细胞系的筛选与鉴定[J].郑州大学学报(医学版),2010,(04):551.
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稳定转染pEGFP4.1N乳癌细胞系的筛选与鉴定()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2010年04期
页码:
551
栏目:
论著
出版日期:
2010-07-30

文章信息/Info

作者:
时晓芳1)闫红霞1)宋安营1)高艳锋1)祁元明1)汲振余2)郭欢1)任岩1)康巧珍1)
1)郑州大学生物工程系 郑州 450001
2)河南省医药科学研究院 郑州 450052
关键词:
乳癌4.1N增强型绿色荧光蛋白真核表达载体MDAMB231
文献标志码:
A
摘要:
目的:建立真核表达载体pEGFP4.1N稳定转染的乳癌MDAMB231细胞系,观察4.1N在MDAMB231细胞中的表达和定位。方法:真核表达载体pEGFP4.1N转化到大肠杆菌中进行扩增,酶切鉴定插入片段后进行测序。脂质体法将重组质粒转染至MDAMB231细胞中,经G418 筛选抗性克隆,利用倒置荧光显微镜观察融合蛋白在细胞中的定位,并用Western blot检测融合蛋白的表达。结果:真核表达载体pEGFP4.1N的酶切片段经琼脂糖电泳和测序鉴定结果正确。G418 筛选14 d后获得pEGFP4.1N稳定转染的MDAMB231抗性克隆。荧光显微镜下观察到融合蛋白在MDAMB231阳性克隆的细胞质中表达,核内不表达。 Western blot亦检测到4.1N融合蛋白的表达。结论:成功建立了pEGFP4.1N稳定转染的乳癌细胞系MDAMB231阳性克隆。

参考文献/References:

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30972707,30873002;国家大学生创新性实验基金资助项目091045922
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