Establishment of a human keratinocyte cell line (HaCaT) stably expressing human papillomavirus type 16 E7 (HPV16E7) protein
10.3760/cma.j.issn.0412-4030.2011.05.006
- VernacularTitle:稳定表达人乳头瘤病毒16E7蛋白HaCaT细胞株的建立
- Author:
Fei MIAO
;
Xiuli WANG
;
Hongwei WANG
;
Huilin DING
;
Ting Lü
;
Linglin ZHANG
- Publication Type:Journal Article
- Keywords:
Human papillomavims 16;
HaCaT cells;
Transfection
- From:
Chinese Journal of Dermatology
2011;44(5):310-313
- CountryChina
- Language:Chinese
-
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.
