| 人红细胞生成素工程细胞株的特性鉴定 |
| |
| 引用本文： | 胡云龙,陈松,徐梅,高坤,林世康,刁振宇,张双全. 人红细胞生成素工程细胞株的特性鉴定[J]. 生物技术, 2004, 14(2): 22-24 |
| |
| 作者姓名： | 胡云龙 陈松 徐梅 高坤 林世康 刁振宇 张双全 |
| |
| 作者单位： | 1. 南京师范大学生命科学院,江苏,南京,210097
2. 南京华欣药业生物工程有限公司,江苏,南京,210002 |
| |
| 摘 要： | 目的：为了对人红细胞生成素工程细胞株(F10—6)进行特性鉴定。方法：将人红细胞生成素基因构建的重组表达质粒pCDB与标志质粒pSV—dhfr共转染CHO-dhfr^-细胞中，经氨甲喋呤加压筛选和亚克隆挑选，获得高效稳定表达EPO的工程细胞株并对其进行鉴定。结果：实验结果表明，工程细胞株无细菌、支原体、真菌和病毒污染；并无致瘤性；染色体数目不变；连续传代50次和三次冻存复苏后，细胞表达稳定。结论：该工程细胞株可用于工业化生产。
|
| 关 键 词： | 人 红细胞生成素 工程细胞株 基因 重组表达质粒 |
| 文章编号： | 1004-311X(2004)02-0022-03 |
| 修稿时间： | 2003-09-30 |
The Analysis and Identify of Characteristics for the Engineering Cell Lines of Erythropoietin |
| |
| HU Yun-long,CHEN Song,XU Mei,GAO Kun,LIN Shi-kang,DIAO Zhen-yu,ZHANG Shuang-quan. The Analysis and Identify of Characteristics for the Engineering Cell Lines of Erythropoietin[J]. Biotechnology, 2004, 14(2): 22-24 |
| |
| Authors: | HU Yun-long CHEN Song XU Mei GAO Kun LIN Shi-kang DIAO Zhen-yu ZHANG Shuang-quan |
| |
| Affiliation: | HU Yun-long~1,CHEN Song~2,XU Mei~2,GAO Kun~2,LIN Shi-kang~2,DIAO Zhen-yu~1,ZHANG Shuang-quan~1 |
| |
| Abstract: | Purpose:The aim was to identify the characteristic of engineering cell of human erythropoietin.Methods:The recombinant plasmid pCDB constructed with EPOcDNA and Marked plasmid pSV-dhfr was cotransfected into CHO-dhfr~- cell.The cloneof the highest expression EPO were selected and identified through methotrexate amplification and subclone.Results:The results of identification show that F10-6 cell line have not been contaminated by mycoplasma,bacteria,fungi,and virus.It also have no the form of tumor in nude mice.The chromosomal number of F10-6 cell line is normal.It was bequeathed for fifty passages and resuscitated after freezing three times.The expression level of EPO is stable.Conclusion:F10-6 engineering cell can be used for the production of industrialization. |
| |
| Keywords: | erythropoietin Engineering cell identification Characteristic |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
