title: [Identification of interaction between BPHL and PML-C].
abstract: OBJECTIVE
  To explore the interaction between BPHL and PML-C by co-immunoprecipitation and yeast two-hybird system.
  
  
  METHODS
  The recombination expression plasmids pGBKT7-PML-C and pACT2-BPHL were cotransformed into yeast AH109, to investigate their interaction in vivo. The expression vector of HA-tagged fusion protein (pCMV-HA-PML-C) and the expression vector of myc-tagged fusion protein (pCMV-myc-BPHL) were constructed and identified respectively, and cotransfected into human embryo kidney 293 (HEK293) cells. The interaction between PML-C and BPHL was investigated by co-immunoprecipitation in vitro.
  
  
  RESULTS
  Blue clones were found in QDO/5-bromo-4-chloro-3-indolyl-alpha-D-galactoside (X-alpha-gal) plate, eukaryotic expression vectors named as pCMV-HA-PML-C and pCMV-myc-BPHL were constructed and confirmed with double restriction enzyme digestion and co-transfected into HEK 293 cells successfully. After immunoprecipitation of HA-PML-C with anti-HA polyclonal antibody, expressed myc-BPHL protein was identified by Western blot with anti-c-myc monoclonal antibody from immunoprecipitated complex.
  
  
  CONCLUSIONS
  The eukaryotic expression vector of PCMV-HA-PML-C and PCMV-myc-BPHL were constructed successfully. The interaction between PML-C and BPHL was identified by co-immunoprecipitation and yeast two-hybird technique.
authors.affiliation: Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Chu
authors.forename: Bei-Zhong
authors.initials: BZ
authors.lastname: Liu
authors.forename: Liang
authors.initials: L
authors.lastname: Zhong
authors.forename: Xiu-Juan
authors.initials: XJ
authors.lastname: Wu
authors.forename: Dan
authors.initials: D
authors.lastname: Zhu
authors.forename: Yanh
authors.initials: Y
authors.lastname: Wu
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D009687:Nuclear Proteins / Q000378:metabolism*; D000071156:Promyelocytic Leukemia Protein; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; C093745:BPHL protein, human; D002265:Carboxylic Ester Hydrolases