title: [Establishment of RAW264.7 cell line stably expressing Mycobacterium tuberculosis protein ESAT-6].
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.
authors: State Key Laboratory ofPathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors: Hao
authors: H
authors: Li
authors: Ying
authors: Y
authors: Yin
authors: Dayong
authors: D
authors: Dong
authors: Jun
authors: J
authors: Zhang
authors: Ling
authors: L
authors: Fu
authors: Chenguang
authors: C
authors: Cai
authors: Meng
authors: M
authors: Wang
authors: Junjie
authors: J
authors: Xu
authors: Wei
authors: W
authors: Chen
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C093276:ESAT-6 protein, Mycobacterium tuberculosis; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins