Spalholz B A, Yang Y C, Howley P M
Cell. 1985 Aug;42(1):183-91. doi: 10.1016/s0092-8674(85)80114-8.
We have mapped a transcriptional regulatory sequence within the 1.0 kb noncoding region of the bovine papilloma virus (BPV-1) genome, using an enhancer dependent expression vector for chloramphenicol acetyltransferase. This transcriptional regulatory element works independently of position and orientation, and its function is significantly augmented in BPV-1 transformed C127 cells and in monkey CV-1 cells acutely transfected with plasmids expressing BPV-1 early gene products. Using defined deletion mutants of the BPV-1 DNA and full-length viral cDNAs expressed from an SV40 early promoter, we demonstrate that the expression of this trans-activating factor maps to the 3' open reading frames of the viral transforming region. A premature termination codon engineered into the E2 ORF eliminates expression of this diffusible transactivation function establishing the E2 gene product as the diffusible trans-activating factor.
我们利用氯霉素乙酰转移酶的增强子依赖性表达载体,绘制了牛乳头瘤病毒(BPV-1)基因组1.0 kb非编码区内的转录调控序列图谱。该转录调控元件的作用与位置和方向无关,并且在BPV-1转化的C127细胞以及急性转染表达BPV-1早期基因产物质粒的猴CV-1细胞中,其功能显著增强。利用BPV-1 DNA的特定缺失突变体和由SV40早期启动子表达的全长病毒cDNA,我们证明这种反式激活因子的表达定位于病毒转化区的3'开放阅读框。在E2开放阅读框中设计的一个提前终止密码子消除了这种可扩散反式激活功能的表达,从而确定E2基因产物为可扩散反式激活因子。