Chinese Hepatolgy ›› 2016, Vol. 21 ›› Issue (1): 34-38.

• Orginal Articles • Previous Articles     Next Articles

Establish ment and expression of recom binant pEGFP-prohibitin plasmid in HepG2 cell line

ZHAI Song, SUN Ming-zhu, WANG Wen-jun, LI Ya-ping, ZHANG Xin, LI Mei, DANG Shuang-suo   

  1. Department of infectious diseases,thesecond affiliated hospital of medicalschool of Xi’an Jiaotong University,Xi’an 710004,China
  • Received:2015-07-07 Published:2020-06-01
  • Contact: DANGShuang-suo,Email:dang212@126.com

Abstract: Objective To construct eukaryotic expression plasmid of enhanced green fluorescent protein plasmids (pEGFP)prohibitin,and to observe its expression in transfected Hep G2 cells. Methods The full coding sequence of prohibitin was am plified by high fidelity poly merase chain reaction(PCR). Mean w hile,the eukaryotic expression vector of pEGFP-prohibitin was constructed and identified by gene sequencing and basic local align ment search tool(BLAST). Plasmids of pEGFP-prohibitin and pEGFP-N1 were extracted and transfected into Hep G2 cells by TurboFect oligofectamine reagent,expression of which in transfected Hep G2 cells was assessed by real time PCRat MRNAlevel and western blot at protein level. Results Recombinant pEGFP-prohibitin plasmid was successfully constructed then transfected into Hep G2 cells,prohibitin expression in which was apparently higher than that in Mock-vehicle and non-transfection groups. Conclusion The eukaryotic expression vector of pEGFP-prohibitin was constructed successfully,and prohibitin was confirmed to be up-regulated in Hep G2/pEGFP-prohibitin cells.

Key words: Prohibitin, Gene expression, Recombinant plasmid, Transfection