Kanda T, Watanabe S, Yoshiike K
Department of Enteroviruses, National Institute of Health, Tokyo, Japan.
Virology. 1988 Jul;165(1):321-5. doi: 10.1016/0042-6822(88)90694-0.
We tested the human papillomavirus 16 (HPV 16) early genes for their ability to immortalize or transform primary rat brain cells, using the expression plasmids that contain the neomycin-resistance-inducing unit (pSV2neo) and the transcriptional unit for the HPV 16 subgenomic DNA fragments controlled by the SV40 early promoter. After transfection, drug-resistant colonies were maintained by refeeding and replating for characterization. The E7 gene alone was found to be capable of immortalizing and morphologically transforming primary rat cells, and the transformed cells showed anchorage-independent growth. Although its activity was lower than that of the E7 gene, the E6 gene also immortalized primary rat cells. The immortalized or transformed cells contained HPV 16-specific DNA and mRNA.
我们使用含有新霉素抗性诱导单位(pSV2neo)以及由SV40早期启动子控制的人乳头瘤病毒16型(HPV 16)亚基因组DNA片段转录单位的表达质粒,测试了HPV 16早期基因使原代大鼠脑细胞永生化或转化的能力。转染后,通过再次喂食和传代培养来维持耐药菌落以进行特性鉴定。结果发现,单独的E7基因能够使原代大鼠细胞永生化并使其发生形态转化,且转化后的细胞呈现出不依赖贴壁的生长特性。尽管E6基因的活性低于E7基因,但它也能使原代大鼠细胞永生化。永生化或转化后的细胞含有HPV 16特异性DNA和mRNA。