Journal of Southern Medical University ›› 2014, Vol. 34 ›› Issue (03): 308-.
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Abstract: Objective To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressingHKE-HERG-G572R. Methods HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated byDNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cellline stably expressing HKE-HERG-G572R. Results The pcDNA3-HERG-G572R expression vector was successfully constructedand the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-foldHKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfectedwith pcDNA3 (P<0.01).Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R toprovide a cell model for studying individualized therapy.
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https://www.j-smu.com/EN/Y2014/V34/I03/308