Stable expression of Mycobacterium tuberculosis protein EspB in RAW264. 7

Abstract:

Abstract:［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 pEGFPC1 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 RTPCR，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．