Journal of Southern Medical University ›› 2014, Vol. 34 ›› Issue (03): 308-.

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Construction of pcDNA3-HERG-G572R expression vector and establishment of a cell line
stably expressing HKE-HERG-G572R

  

  • Online:2014-03-20 Published:2014-03-20

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).Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to
provide a cell model for studying individualized therapy.