Thummel C, Tjian R, Grodzicker T
Cell. 1981 Mar;23(3):825-36. doi: 10.1016/0092-8674(81)90447-5.
We have obtained novel adenovirus-SV40 recombinant viruses that express wild-type SV40 large T and small t antigens under the control of different adenovirus promoters. Hybrids were constructed in vitro with SV40 DNA that contains the entire early coding region but lacks the transcriptional promoter. Recombinants were isolated by a strong biological selection for viruses that express SV40 T antigen. Analysis of several recombinant genomes indicates that they contain the SV40 A gene inserted in a variety of positions and orientations in the adenoviral genome. Moreover, the set of hybrid transcripts reveals an unexpected variety of splicing patterns. Some hybrid mRNAs transcribed from the adenovirus late promoter appear to contain the adenovirus tripartite leader sequence. Other hybrid mRNAs were transcribed from adenovirus early promoters. All recombinant mRNAs contain intact SV40 early sequences that have normal splice patterns and produce wild-type T antigens. Biochemical characterization of SV40 T antigens overproduced by the hybrid viruses indicates that they are structurally indistinguishable from wild-type SV40 large T antigen and are functionally equivalent to the D2 protein.
我们获得了新型腺病毒 - SV40重组病毒，这些病毒在不同腺病毒启动子的控制下表达野生型SV40大T抗原和小t抗原。用含有完整早期编码区但缺乏转录启动子的SV40 DNA在体外构建杂种。通过对表达SV40 T抗原的病毒进行强大的生物学筛选来分离重组体。对几个重组基因组的分析表明，它们含有插入腺病毒基因组中各种位置和方向的SV40 A基因。此外，杂种转录本的集合揭示了意想不到的多种剪接模式。一些从腺病毒晚期启动子转录的杂种mRNA似乎含有腺病毒三联前导序列。其他杂种mRNA则从腺病毒早期启动子转录。所有重组mRNA都含有完整的SV40早期序列，这些序列具有正常的剪接模式并产生野生型T抗原。杂种病毒过量产生的SV40 T抗原的生化特性表明，它们在结构上与野生型SV40大T抗原无法区分，并且在功能上等同于D2蛋白。
