title: [Construction, expression and cell growth inhibition of translocating peptide/granzyme B fusion protein gene].
abstract: OBJECTIVE
  To investigate the inhibitory effect of translocating peptide/granzyme B fusion protein on cell growth.
  
  
  METHODS
  The translocating peptide gene of Pseudomonas exotoxin A (PE) was fused with active granzyme B gene by recombinant PCR to construct PE II-GrBa fusion protein gene. PE II-mGrBa with a mutation of serine to cystein at active center of GrB was used as negative control. The resulting PE II-GrBa and PE II-mGrBa genes were transiently transfected into mammalian cells via lipofectamine mediation. The effects of expression of PE II-GrBa gene on morphology and growth of transfected cells were detected by MTT colorimetry, TUNEL assay and indirect immunofluorescence staining.
  
  
  RESULTS
  Transient expression of PE II-GrBa resulted in cytoskeleton abnormality, cell growth inhibition, and apoptosis in some cells.
  
  
  CONCLUSIONS
  Expression of PE II-GrBa fusion protein can inhibit cell growth.
authors.affiliation: Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032 China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Zhi
authors.initials: Z
authors.lastname: Wang
authors.forename: Yan-ming
authors.initials: YM
authors.lastname: Xu
authors.forename: Jun-hao
authors.initials: JH
authors.lastname: Gui
authors.forename: Wei-hong
authors.initials: WH
authors.lastname: Wen
authors.forename: Cheng-ji
authors.initials: CJ
authors.lastname: Wang
authors.forename: An-gang
authors.initials: AG
authors.lastname: Yang
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D005098:Exotoxins / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D053804:Granzymes; D006367:HeLa Cells; D006801:Humans; D010446:Peptide Fragments / Q000235:genetics; D011549:Pseudomonas; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
chemical_list: D005098:Exotoxins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases