China Animal Husbandry and Veterinary Medicine ›› 2020, Vol. 47 ›› Issue (5): 1352-1359.doi: 10.16431/j.cnki.1671-7236.2020.05.008

• Physiology and Biochemistry • Previous Articles     Next Articles

Comparative Study on Three Methods of Mediated Porcine NR2F2 Gene Transfection into PK15 Cells

ZHANG Wanfeng, HEI Wei, HE Zhiqiang, LIU Min, MENG Shan, GAO Pengfei, CAI Chunbo, YANG Yang, GUO Xiaohong, CAO Guoqing, LI Bugao   

  1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China
  • Online:2020-05-20 Published:2020-05-18

Abstract: The aim of this study was to compare the efficiency of transfected pig kidney epithelial cells (PK15) by three transfected methods,so as to lay the foundation for studying the function of foreign genes with PK15 cells as a model.In this study,PK15 cells as research subject were transfected with liposome,electroporation and lentivirus methods.The transfected efficiency was evaluated based on the expression of green fluorescent protein observed by fluorescence microscope,the expressions of EGFP and NR2F2 genes were detected by Real-time quantitative PCR and the cell survival rate was detected by CCK-8 reagent.The results showed that the transfected efficiency of PK15 cells transfected with electroporation and lentivirus methods was extremely significantly higher than that of liposomes (P<0.01),whereas the difference between electroporation and lentivirus methods was not significant (P>0.05).The Real-time quantitative PCR results showed that the expressions of EGFP and NR2F2 genes in PK15 cell transfected with lentivirus method were the highest and the lowest with liposome method,which was in accordance with the transfected efficiency.CCK-8 results showed that the cell survival rate transfected by electransfection was the lowest,which was extremely significantly lower than that of control group (P<0.01).The cell survival rate transfected with the liposome was significantly lower than that of control group (P<0.05).The difference of cell survival rate between lentivirus and control group was not significant (P>0.05).Based on the transfection efficiency and cell viability of this study,the lentiviral transfection method was the suitable way for transfection of PK15 cells,which provided a perfect method for high-efficiency transfection of PK15 cells in the future.

Key words: PK15 cells; NR2F2 gene; liposome transfection; electroporated transfection; lentiviral transfection

CLC Number: