title: [Construction of eukaryotic expression vector of F-box protein 6 gene and its expression in HEK293T cells].
abstract: OBJECTIVE
  To construct eukaryotic expression vector of F-box protein 6 (FBXO6) gene.
  
  
  METHODS
  The full-length FBXO6 cDNA containing two restriction sites (BglII and EcoRI) was synthesized by PCR. The pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 vectors were constructed respectively. The size and sequence of cDNA fragments were confirmed by bacterial colony PCR and BglII and EcoRI digestion and sequencing. HEK293T cells were transfected with the pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 vectors respectively, and then the expression of FBXO6 protein was detected using Western blotting.
  
  
  RESULTS
  The pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 contained FBXO6 fragments of proper size and sequence. FBXO6 protein was expressed efficiently in pEGFP-C1-FBXO6 transfected HEK293T cells, but it was down-regulated in pEGFP-C1-anti-FBXO6 transfected 293T cells.
  
  
  CONCLUSIONS
  Both pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 were successfully constructed.
authors.affiliation: Institute of Genetic Engineering, Southern Medical University, Guangzhou, China. gaoyuan1623@163.com
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gao
authors.forename: Jinghua
authors.initials: J
authors.lastname: Feng
authors.forename: Gan
authors.initials: G
authors.lastname: Chen
authors.forename: Yifei
authors.initials: Y
authors.lastname: Peng
authors.forename: Wenling
authors.initials: W
authors.lastname: Zheng
authors.forename: Wenli
authors.initials: W
authors.lastname: Ma
mesh_terms: D000906:Antibodies / Q000235:genetics; D003001:Cloning, Molecular; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis*; D044843:SKP Cullin F-Box Protein Ligases / Q000235:genetics* / Q000276:immunology; D014162:Transfection
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C479752:FBXO6 protein, human; D044843:SKP Cullin F-Box Protein Ligases