title: [Study of localization and translocation of human high mobility group protein B1 in eukaryotic cells].
abstract: OBJECTIVE
  To observe the localization and translocation of human high mobility group protein B1 (HMGB1) protein in eukaryotic cells.
  
  
  METHODS
  The vector that expressed the fusion protein of human HMGB1 and enhanced green fluorescent protein (EGFP) in mammalian cells were constructed. HMGB1 and EGFP were subcloned into hytohemagglutinin (HA) tagged vector pcDNA3-HA by two steps, the construct of which was then transfected into 293A cells and observed with fluorescence microscope.
  
  
  RESULTS
  The recombinant plasmid was verified by enzyme digestion, polymerase chain reaction (PCR) and sequence analysis, and the fusion protein was highly expressed in 293A cells. With fluorescence microscopy the green fluorescence was found to be distributed in the nuclei. Eighteen hours after the stimulation with tumor necrosis factor-alpha (TNF-alpha), it was found to have translocated into the cytoplasm in some of the cells.
  
  
  CONCLUSIONS
  The expression vector for HMGB1-EGFP fusion protein is successfully constructed and effectively expressed in mammalian cells, and it can serve as an important tool in the study of the signal pathway of HMGB1 in mammalian cells.
authors.affiliation: Department of Pathophysiology and Key Laboratory of Functional Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, Guangdong, China. graceXjj@126.com
authors.forename: Jia
authors.initials: J
authors.lastname: Xu
authors.forename: Zhi-feng
authors.initials: ZF
authors.lastname: Liu
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Deng
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel; D005057:Eukaryotic Cells / Q000378:metabolism; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D024243:HMGB1 Protein / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D011995:Recombination, Genetic
chemical_list: D024243:HMGB1 Protein; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins