| 正交设计优化电转染CHO细胞的方法 |
| |
| 引用本文： | 陆俭,李萍,马亚茹,陈勇,蒋琳.正交设计优化电转染CHO细胞的方法[J].生物技术通讯,2011,22(5):713-716. |
| |
| 作者姓名： | 陆俭 李萍 马亚茹 陈勇 蒋琳 |
| |
| 作者单位： | 兰州生物制品研究所,甘肃兰州,730046 |
| |
| 摘 要： | 目的:探讨细胞电穿孔转染的优化方法。方法:按照L9(34)正交表安排各因素水平的细胞电敏感性实验,以台盼蓝染色计数确定细胞存活率,最接近50%存活率为最优方案,根据最优方案将CHO-dhfr-细胞与DNA混匀电穿孔转染以验证正交设计结果。结果:CHO-dhfr-细胞电转染的优化参数为低离子强度Tris-Cl缓冲液、750 V/cm、50μF、电击2次、间隔1 min,各因素对转染的影响大小依次为缓冲液、电场强度、脉冲次数、电容。结论:确定了电穿孔法转染CHO-dhfr-细胞的方法,为进一步应用该细胞株表达重组蛋白打下了基础。
|
| 关 键 词： | 电穿孔法 CHO-dhfr-细胞 正交设计 |
Study on Optimizing the Method of Electrotransfection Applying CHO cell by Orthogonal Design |
| |
| LU Jian,LI Ping,MA Ya-Ru,CHEN Yong,JIANG Lin.Study on Optimizing the Method of Electrotransfection Applying CHO cell by Orthogonal Design[J].Letters in Biotechnology,2011,22(5):713-716. |
| |
| Authors: | LU Jian LI Ping MA Ya-Ru CHEN Yong JIANG Lin |
| |
| Institution: | LU Jian,LI Ping,MA Ya-Ru,CHEN Yong,JIANG Lin Lanzhou Institute of Biological Products,Lanzhou 730046,China |
| |
| Abstract: | Objective: To explore the optimized method of electrotransfection.Methods: Design the cell electro-susceptibility experiment of the factors and levels according to L 9(3 4) orthogonal layout.Confirm the cell livability by cell counting of trypan blue staining.The best project is the fact that cell livability nearing 50%.Perform the electrotransfection according to the best project in order to validate the result of orthogonal design.Results: The optimized parameter of electrotrans-fection for CHO-dhfr-cells... |
| |
| Keywords: | electrotransfection CHO-dhfr-cell orthogonal design |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
