Establishment of RAW264.7 cell line stably expressing Mycobacterium tuberculosis protein ESAT-6.
- Author:
Hao LI
1
;
Ying YIN
;
Dayong DONG
;
Jun ZHANG
;
Ling FU
;
Chenguang CAI
;
Meng WANG
;
Junjie XU
;
Wei CHEN
Author Information
1. State Key Laboratory ofPathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antigens, Bacterial;
biosynthesis;
genetics;
Bacterial Proteins;
biosynthesis;
genetics;
Cell Line;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Macrophages;
cytology;
metabolism;
Mice;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Transfection
- From:
Chinese Journal of Biotechnology
2011;27(9):1390-1396
- CountryChina
- Language:Chinese
-
Abstract:
For studying the effects of Mycobacterium tuberculosis secretory protein ESAT-6 on the related functions of macrophages, RAW264.7 cells were transfected with pEGFP-C1-ESAT-6 and pEGFP-C1 by liposome respectively. After screening with a high level of G418, the macrophage cell lines that stably expressed EGFP-ESAT-6 fusion protein or EGFP were established. The gene and protein expression levels were further analyzed by RT-PCR, fluorescence microscopy and Western blotting. The results indicated that the EGFP-ESAT6 fusion gene was integrated into the chromosome and the protein could be stably expressed in the selected macrophage cell line. These results gave us a tool for the future study in the mechanisms of ESAT-6 protein in modulating the macrophage cells.
