Efficient transactivation and morphologic transformation by bovine papillomavirus genes expressed from a bovine papillomavirus/simian virus 40 recombinant virus.
Efficient transactivation and morphologic transformation by bovine papillomavirus genes expressed from a bovine papillomavirus/simian virus 40 recombinant virus.
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牛乳头瘤病毒/猿猴病毒 40 重组病毒表达的牛乳头瘤病毒基因的高效反式激活和形态转化。
DOI：
10.1073/pnas.85.23.9007
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发表时间：
1988
影响因子：
11.1
通讯作者：
DiMaio,D
中科院分区：
文献类型：
--
作者：
Settleman,J;DiMaio,D
为了有效地将牛乳头瘤病毒1型基因导入培养的细胞中，我们构建了一个杂交病毒基因组，其中猴病毒40早期区域被替换为牛乳头瘤病毒1型转化区的片段。在表达猴病毒40大肿瘤抗原的猴细胞中产生含有重组基因组的猴病毒40病毒粒子的高滴度原液。用该病毒感染的细胞有效地表达牛乳头瘤病毒1型E2和E5基因。E2基因的表达引起与牛乳头瘤病毒1型控制区相关的基因的反式激活，导致高达1000倍的诱导。在含有整合报告基因的细胞系的高感染复数下，大多数细胞被感染并对反式激活作出反应。在用野生型病毒感染而不是用开放阅读框E5突变体感染的48小时内，小鼠C127细胞显示出与致瘤转化中所见相似的形态和生长特征的显著变化。该系统可用于确定对牛乳头瘤病毒1型转化和调节基因的引入的急性细胞反应;它也可用于诱导外源基因稳定地掺入培养的哺乳动物细胞中。
To efficiently introduce bovine papillomavirus type 1 genes into cultured cells, we constructed a hybrid viral genome in which the simian virus 40 early region is replaced with a segment of the bovine papillomavirus type 1 transforming region. High-titer stocks of simian virus 40 virions containing the recombinant genome were produced in monkey cells that express simian virus 40 large tumor antigen. Cells infected with this virus efficiently expressed the bovine papillomavirus type 1 E2 and E5 genes. Expression of the E2 gene caused transactivation of genes linked to the bovine papillomavirus type 1 control region, resulting in up to a 1000-fold induction. At high multiplicity of infection of a cell line containing an integrated reporter gene, most cells were infected and responded to transactivation. Within 48 hr of infection with wild-type virus but not with an open reading frame E5 mutant, mouse C127 cells displayed dramatic changes in morphology and growth characteristics similar to those seen in tumorigenic transformation. This system can be used to determine the acute cellular response to introduction of bovine papillomavirus type 1 transforming and regulatory genes; it can also be used to induce foreign genes stably incorporated into cultured mammalian cells.