| 阳离子脂质体转染家蚕培养细胞的技术体系研究 |
| |
| 引用本文： | 潘敏慧,刘敏,刘佳,杜娟,张春东,鲁成. 阳离子脂质体转染家蚕培养细胞的技术体系研究[J]. 蚕业科学, 2008, 34(4) |
| |
| 作者姓名： | 潘敏慧 刘敏 刘佳 杜娟 张春东 鲁成 |
| |
| 作者单位： | 西南大学,农业部蚕桑学重点开放实验室,重庆,400716;西南大学,农业部蚕桑学重点开放实验室,重庆,400716;云南省农业科学院蚕桑蜜蜂研究所,云南蒙自,661101 |
| |
| 基金项目： | 国家重点基础研究发展规划(973计划)，国家高技术研究发展计划(863计划)，国家自然科学基金 |
| |
| 摘 要： | 阳离子脂质体转染家蚕培养细胞是家蚕基因功能研究的重要技术之一。利用阳离子脂质体Lipofectamine2000与pBac[A3Neo+A3EGFP]转基因表达载体,对家蚕卵巢细胞BmN-SWU1、BmN-SWU2、BmN和草地贪夜蛾卵巢细胞Sf21进行了外源DNA质量、脂质体用量、转染细胞密度和转染孵育时间等4个转染条件的优化实验,筛选到了阳离子脂质体转染效率最高的细胞系BmN-SWU1。BmN-SWU1细胞的最佳转染条件为:外源DNA0.6μg,脂质体2.5μL,细胞密度1×105mL-1,无血清条件下孵育24 h。此条件下转染效率可达55.08%。用抗生素G418筛选转染后的BmN-SWU1细胞,建立了GFP转基因细胞系。
|
| 关 键 词： | 家蚕 阳离子脂质体 细胞转染 转基因细胞系 |
Technical System of Silkworm Cell Lines Transfected With Cation Liposome |
| |
| PAN Min-Hui,LIU Min,LIU Jia,DU Juan,ZHANG Chun-Dong,LU Cheng. Technical System of Silkworm Cell Lines Transfected With Cation Liposome[J]. Acta Sericologica Sinica, 2008, 34(4) |
| |
| Authors: | PAN Min-Hui LIU Min LIU Jia DU Juan ZHANG Chun-Dong LU Cheng |
| |
| Abstract: | It is an important technique for studying on functional genes of silkworm that silkworm cell lines were transfected with cation liposome.In this paper,4 factors of transfection of pBac[A3Neo+A3EGFP] with Lipofectamine 2000 in 4 insect cell lines,including the DNA amount,liposome amount,cells density and transfection duration,were investigated and optimized.The results showed that the transfection efficiency of BmN-SWU1 was 55.08%,highest in 4 insect cell lines tested.The optimal transfection conditions for BmN-SWU1 were as following: 0.6 μg DNA,2.5 μL cation liposome,1×105 cells/mL and transfection duration 24 h.BmN-SWU1 was transfected with pBac[A3Neo+A3EGFP],then selected under G418 stress.Finally,a cell line with over 95% transgenic cells was obtained. |
| |
| Keywords: | Bombyx mori Cation liposome Cell transfection Transgenic cell line |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
