首页> 美国卫生研究院文献>Journal of Virology >Simian virus 40 late promoter region able to initiate simian virus 40 early gene transcription in the absence of the simian virus 40 origin sequence.
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Simian virus 40 late promoter region able to initiate simian virus 40 early gene transcription in the absence of the simian virus 40 origin sequence.

机译:在没有猿猴病毒40起源序列的情况下能够启动猿猴病毒40早期基因转录的猿猴病毒40晚期启动子区域。

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摘要

To improve our knowledge of the simian virus 40 (SV40) late promoter control region, we took advantage of the fact that T antigen can be expressed with a heterologous promoter. We constructed three chimeric plasmids (pEMP-273, pEMP-LCAP-162, and pEMP-LCAP-113) each with a putative late promoter sequence positioned immediately upstream from the SV40 early gene coding region but in an orientation opposite to its natural orientation in the SV40 genome. After transfection of the recombinant DNA into HeLa or CV1 cells, T antigen accumulation, as scored by indirect immunofluorescence, was used as a functional test for promoter activity. We found that the sequence mapping from nucleotides 332 to 273 is not sufficient for promoting transcription of SV40 early gene but does potentiate the promoter activity of the 72-base-pair repeats in initiating the transcription at natural late cap sites. Considering that both plasmids pEMP-LCAP-162 and pEMP-LCAP-113 lack all of the sequence of the SV40 replication origin, we conclude that SV40 transcription can be mediated through a putative late promoter in the absence of the sequence for the SV40 replication origin.
机译:为了提高我们对猿猴病毒40(SV40)晚期启动子控制区的了解,我们利用了T抗原可以与异源启动子一起表达这一事实。我们构建了三个嵌合质粒(pEMP-273,pEMP-LCAP-162和pEMP-LCAP-113),每个质粒均具有一个推定的晚期启动子序列,该序列位于SV40早期基因编码区的紧邻上游,但方向与其天然方向相反。 SV40基因组。在将重组DNA转染到HeLa或CV1细胞中后,通过间接免疫荧光评分将T抗原蓄积用作启动子活性的功能测试。我们发现从核苷酸332到273的序列映射不足以促进SV40早期基因的转录,但确实增强了72个碱基对重复序列的启动子活性,从而在自然的晚期帽位启动了转录。考虑到质粒pEMP-LCAP-162和pEMP-LCAP-113都缺少SV40复制起点的所有序列,我们得出结论,在不存在SV40复制起点的序列的情况下,可以通过推定的晚期启动子介导SV40转录。 。

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