title: [Stable expression of Mycobacterium tuberculosis protein EspB in RAW264.7].
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 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.
  
  
  CONCLUSIONS
  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.
authors: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China. boyleehao@126.com
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: Jun
authors: J
authors: Ren
authors: Junjie
authors: J
authors: Xu
authors: Wei
authors: W
authors: Chen
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000235:genetics; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins