2011•Chinese Journal of DermatologyRequires access
Establishment of a humankeratinocyte cell line (HaCaT) stably expressing human papillomavirus type 16 E7 (HPV16E7)protein
Fei Miao, Xiuli Wang, Hongwei Wang, Huilin Ding, Lei Ting, Linglin Zhang
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Abstract
：Objective To establish ahuman keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein. Methods HPV16E7gene was amplified from CaSki cells using PCR and inserted into the eukaryotic expressionplasmid pcDNA3.1. Then, the recombinant expression plasmid pcDNA3.1-HPV16E7 wastransfected into HaCaT cells followed by G418 selection and identification by RT-PCR andWestern blot. Results The recombinant eukaryotic expression plasmid pcDNA3.1-HPV16E7 wassuccessfully identified by restriction enzyme digestion pattern and sequence analysis.Agarose gel electrophoresis of RT-PCR products detected the 297-bp fragment of HPV16E7cDNA, and Western blot confirmed the stable expression of HPV16E7 protein. Conclusion Ahuman keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein is successfullyestablished.