title: [Construction of pcDNA3-HERG-G572R expression vector and establishment of a cell line stably expressing HKE-HERG-G572R].
abstract: OBJECTIVE
  To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R.
  
  
  METHODS
  HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R.
  
  
  RESULTS
  The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).
  
  
  CONCLUSIONS
  The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.
authors.affiliation: Department of Genetics and Molecular Biology, Xi'an Jiaotong University College of Medicine, Xi'an 710061, China. E-mail: yysmile_ok@stu.xjtu.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Na
authors.initials: N
authors.lastname: Huang
authors.forename: Ling
authors.initials: L
authors.lastname: Gao
authors.forename: Su'e
authors.initials: S
authors.lastname: Chang
authors.forename: Bo
authors.initials: B
authors.lastname: Guo
authors.forename: Lili
authors.initials: L
authors.lastname: Hu
authors.forename: Tusheng
authors.initials: T
authors.lastname: Song
authors.forename: Chen
authors.initials: C
authors.lastname: Huang
mesh_terms: D001483:Base Sequence; D051638:Ether-A-Go-Go Potassium Channels / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D057809:HEK293 Cells / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D014162:Transfection
chemical_list: D051638:Ether-A-Go-Go Potassium Channels