人类乳头瘤病毒16全基因在体外培养的正常人角质形成细胞中的瞬时表达

摘要/Abstract

摘要： 目的利用含有人类乳头瘤病毒(HPV)16全基因的质粒转染正常人角质形成细胞,观察HPV16 mRNA在转染细胞的表达。方法用FuGENE^TM6转染试剂,将携带HPV16全基因的质粒pSV2-neo/16转染体外培养的正常人角质形成细胞,在转染后24h提取细胞总RNA和DNA,进行RT-PCR和Southern印迹分析。结果 24h后,RT-PCR成功地扩增出110bp的片段,转染细胞中已出现HPV16 mRNA的表达,Southern印迹证实转染细胞中含有HPV16全基因7.9kb。结论 pSV2-neo/16转染正常人表皮角质形成细胞,24h内即可检测到HPV16mRNA表达。

关键词: 乳头状瘤病毒,人, 质粒, 角蛋白细胞, 转染

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

左亚刚, 王家璧, 王宝玺. 人类乳头瘤病毒16全基因在体外培养的正常人角质形成细胞中的瞬时表达[J]. 中华皮肤科杂志, 2000, 33(4): 266-268.doi:

ZUO Yagang, WANG Jiabi, WANG Baoxi. Transient Expression of a Transfected HPV16 Whole Gene in Normal Human Keratinocytes[J]. Chinese Journal of Dermatology, 2000, 33(4): 266-268.doi:

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