| 结核分枝杆菌HSP65-hIL-2融合基因稳定转染细胞系的建立 |
| |
| 引用本文： | 王丽梅,徐志凯,柏银兰,张薇,康健,师长宏.结核分枝杆菌HSP65-hIL-2融合基因稳定转染细胞系的建立[J].科学技术与工程,2009,9(11). |
| |
| 作者姓名： | 王丽梅 徐志凯 柏银兰 张薇 康健 师长宏 |
| |
| 作者单位： | 1. 第四军医大学基础部微生物学教研室,西安,710032
2. 第四军医大学实验动物中心,西安,710032 |
| |
| 摘 要： | 建立可稳定表达结核分枝杆菌HSP65-hIL-2融合蛋白的稳定转染P815细胞系. 在阳离子脂质体作用下,将HSP65-hIL-2真核表达质粒转染与BALB/c 遗传背景一致的P815细胞(H-2d).G418筛选阳性克隆,RT-PCR和间接免疫荧光法检测目的蛋白的转录和表达. 阳性克隆细胞经RT-PCR检测到HSP65-hIL-2融合基因特异性的mRNA表达;用鼠抗人的IL-2 mAb进行间接免疫荧光检测,可在转染的P815细胞浆中观察到较强的绿色特异性荧光,而未转染细胞则为阴性.成功获得稳定表达HSP65-hIL-2融合蛋白的稳定转染细胞系,为其疫苗的CTL研究提供了合适的靶细胞.
|
| 关 键 词： | 结核分枝杆菌 稳定转染 |
Establishment of a Eell Line Stably Transfected the Fusion Gene HSP65-hIL-2 of Mycobacterium tuberculosis |
| |
| WANG Li-mei,XU Zhi-kai,BAI Yin-lan,ZHANG Wei,KANG Jian,SHI Chang-hong.Establishment of a Eell Line Stably Transfected the Fusion Gene HSP65-hIL-2 of Mycobacterium tuberculosis[J].Science Technology and Engineering,2009,9(11). |
| |
| Authors: | WANG Li-mei XU Zhi-kai BAI Yin-lan ZHANG Wei KANG Jian SHI Chang-hong |
| |
| Institution: | Department of Microbiology;School of Basic Medicine;Laborabory Animal Research Center1;Fourth Military Medical University;Xi'an 710032;P.R.China |
| |
| Abstract: | To establish the stably transfected P815 cell line expressing the fusion protein HSP65-hIL-2,the eukaryotic expression plasmid pcDNA-HSP65-hIL-2 was stably transfected into P815 cells(H-2d) using the cationic liposome,whose genetic background was identical to that of BALB/c mice.The positive transfected cells were selected by G418,the specific mRNA of HSP65-hIL-2 was detected by RT-PCR and the expression of the fusion protein in the transfected cells was detected by indirect immunofluorescence technique.The... |
| |
| Keywords: | HSP65 hIL-2 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
