Mansour S L, Grodzicker T, Tjian R
Proc Natl Acad Sci U S A. 1985 Mar;82(5):1359-63. doi: 10.1073/pnas.82.5.1359.
We have used a generalized adenovirus vector system to express the three polyoma tumor (T) antigen proteins under the control of the adenovirus major late promoter. One hybrid virus, Ad-PySVR498, expresses high levels of polyoma middle and small T antigens. A second hybrid virus, Ad-LTSVR545, which contains a cDNA copy of the polyoma A gene, overproduces large T antigen. The T antigens produced are indistinguishable from their authentic polyoma counterparts as determined by immunoprecipitation and partial cleavage by V8 protease. Analysis of polyoma mRNAs encoded by the recombinant viruses showed that they initiate from the adenovirus major late promoter and contain the tripartite leader at their 5' ends. Large T antigen isolated from Ad-LTSVR545-infected cells by immunoaffinity was shown to bind selectively to polyoma DNA sequences that contain the origin of viral DNA replication as well as the sites for transcription initiation.
我们使用了一种通用腺病毒载体系统,在腺病毒主要晚期启动子的控制下表达三种多瘤病毒肿瘤(T)抗原蛋白。一种杂交病毒Ad-PySVR498可高水平表达多瘤病毒中T抗原和小T抗原。第二种杂交病毒Ad-LTSVR545含有多瘤病毒A基因的cDNA拷贝,可过量产生大T抗原。通过免疫沉淀和V8蛋白酶部分切割确定,所产生的T抗原与其天然多瘤病毒对应物无法区分。对重组病毒编码的多瘤病毒mRNA的分析表明,它们从腺病毒主要晚期启动子起始,并且在其5'端含有三联前导序列。通过免疫亲和从Ad-LTSVR545感染的细胞中分离出的大T抗原显示,其可选择性结合包含病毒DNA复制起点以及转录起始位点的多瘤病毒DNA序列。