Chinese Journal of Dermatology ›› 2011, Vol. 44 ›› Issue (5): 310-313.

• Original articles • Previous Articles     Next Articles

Establishment of a human keratinocyte cell line （HaCaT） stably expressing human papillomavirus type 16 E7 （HPV16E7） protein

  

  • Received:2010-07-01 Revised:2010-08-02 Online:2011-05-15 Published:2011-05-03
  • Supported by:

    Study of Apoptotic Mechanism Induced by 5-Aminolevulinic Acid-Photodynamic Therapy in Keratinocytes of HPV16E7 Transfection

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Abstract:

Objective To establish a human keratinocyte cell line （HaCaT） stably expressing HPV16E7 protein. Methods HPV16E7 gene was amplified from CaSki cells using PCR and inserted into the eukaryotic expression plasmid pcDNA3.1. Then, the recombinant expression plasmid pcDNA3.1-HPV16E7 was transfected into HaCaT cells followed by G418 selection and identification by RT-PCR and Western blot. Results The recombinant eukaryotic expression plasmid pcDNA3.1-HPV16E7 was successfully identified by restriction enzyme digestion pattern and sequence analysis. Agarose gel electrophoresis of RT-PCR products detected the 297-bp fragment of HPV16E7 cDNA, and Western blot confirmed the stable expression of HPV16E7 protein. Conclusion A human keratinocyte cell line （HaCaT） stably expressing HPV16E7 protein is successfully established.

Key words: Stable cell lines