Transient Expression of Human Papillomavirus Type 16(HPV 16) mRNA in Normal Human Keratinocytes Tran

• 作 者: 左亚刚 王家璧 王宝玺
• 机 构: Department of Dermatology Peking Union Medical College Hospital,Department of Dermatology,Peking Uni
• 出 处: Chinese Journal of Sexually Transmitted Infections
• 发表日期: 2002年02期
• 关键词: DNA Plasmid plasmid cultured transfection BamHI reagent RNA agarose maintain

Objective: To investigate the expression of HPV16 mRNA innormal human keratinocytes transfected with pSV2-neo/16. Methods: First human keratinocytes were cultured in theserum-free medium M154.Second, the plasmid pSV2-neo/16was transfected into the human keratinocytes using atransfecting reagent. Third, RT-PCR and Southern Blottingwere used to detect the expression of HPV16 mRNA and DNAin the transfected keratinocytes, respectively. Results: The expression of HPV 16 mRNA was successfullyamplified and an 110bp was detected by RT-PCR. A 7.9kbfragment was confirmed in the transfected keratinocytes bySouthern Blot analysis. Conclusion: HPV 16 mRNA and DNA were successfullydetected in the human keratinocytes.

Objective: To investigate the expression of HPV16 mRNA innormal human keratinocytes transfected with pSV2-neo / 16. Methods: First human keratinocytes were cultured in the medium-free medium M154.Second, the plasmid pSV2-neo / 16 was transfected into the human keratinocytes using atransfecting Third, RT-PCR and Southern Blotting used to detect the expression of HPV16 mRNA and DNA in the transfected keratinocytes, respectively. Results: The expression of HPV 16 mRNA was successfully amplified and 110 bp was detected by RT-PCR. A 7.9 kbfragment was confirmed in the transfected keratinocytes by Southern Blot analysis. Conclusion: HPV 16 mRNA and DNA were successfully detected in the human keratinocytes.