Abstract: Objective To investigate the expression of HPV16 mRNA in normal human keratinocytes transfected with a plasmid containing HPV16 whole gene. Methods Normal human keratinocytes were cultured in vitro with HPV16 whole gene and FuGENE^TM 6, a transfecting reagent, then total RNA and DNA were extracted. RT-PCR and Southern blot were used to identify the expression of HPV16 mRNA and DNA, respectively. Results Tweenty four hours after transfection HPV mRNA was expressed in transfected human keratinocytes and a 110 bp product was successfully amplified by RT-PCR method.A 7.9 kb DNA fragment, which corresponded to HPV16 whole gene, was confirmed in the transfected keratinocytes by Southern blot analysis. Conclusion Tweenty four hours after transfection of normal human keratinocytes with pSV2-neo/16,the expression of HPV16 mRNA and DNA was successfully detected.

Key words: Papillomavirus,human, Plasmids, Keratinocytes, Transfection