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Updated: May 28, 2026
Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria
Published on: December 8, 2023
[Stable expression of Mycobacterium tuberculosis protein EspB in RAW264.7]
Hao Li1, Ying Yin, Dayong Dong
1State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China. boyleehao@126.com
Objective:
We established a cell line stably expressing Mycobacterium tuberculosis secretory protein EspB in order to provide evidences for studying EspB in modulating the functions of macrophage.
Methods:
The recombinant plasmid pEGFP-C1-EspB was first constructed, then RAW264.7 cell was transfected with pEGFP-C1-EspB and pEGFP-C1 by liposome respectively. After screening with a high level of G418, the macrophage cell lines that stably expressed EGFP-EspB fusion protein or EGFP were established. The gene and protein expression levels were further analyzed by RT-PCR, fluorescence microscopy and western blot.
Results:
The EGFP-EspB fusion gene was integrated into the chromosome and the protein was stably expressed in the selected macrophage cell line. The macrophage cell lines that stably expressed EGFP-EspB fusion protein or EGFP were established.
Conclusion:
These results gave us a tool for the future study in the effects of EspB protein in modulating the functions of macrophage and its interaction with other molecules of macrophage.
