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首页> 外文期刊>Molecular and Cellular Biology >Functional organization of the simian virus 40 origin of DNA replication.
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Functional organization of the simian virus 40 origin of DNA replication.

机译：猿猴病毒的功能组织40 DNA复制的起源。

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摘要

To define the sequence elements involved in initiation of DNA synthesis at the simian virus 40 origin of replication, we determined the relative replication efficiencies in vitro and in vivo of templates containing a variety of mutations within the origin region. Replication of the mutants in vitro was assayed by the cell-free DNA replication system that we recently described (J.J. Li and T.J. Kelly, Proc. Natl. Acad. Sci. USA 81:6973-6977, 1984; J.J. Li and T.J. Kelly, Mol. Cell. Biol. 5:1238-1246, 1985), and replication in vivo was assayed after transfection of the mutant templates into COS-1 cells. The minimal origin of replication defined by both assays included a 15-base-pair (bp) imperfect inverted repeat, a 27-bp perfect inverted repeat, and a 17-bp A/T-rich region. T-antigen binding site I was not required for DNA replication, but its presence increased replication efficiency severalfold both in vitro and in vivo. Although SP1 binding sites and enhancers had little or no effect on replication in vitro, the presence of either element markedly increased replication in vivo. Thus, the biological role of these elements is not restricted to stimulating transcription but may be more general.
机译：为了定义在猿猴病毒40复制起点中启动DNA合成的序列元素，我们确定了在起始区域内包含各种突变的模板的体外和体内相对复制效率。通过我们最近描述的无细胞DNA复制系统对突变体的体外复制进行了检测（JJ Li和TJ Kelly，美国国家科学院院刊81：6973-6977，1984; JJ Li和TJ Kelly， （Mol.Cell.Biol.5：1238-1246，1985），并在将突变体模板转染至COS-1细胞后测定了体内复制。两种测定法定义的最小复制起点包括一个15个碱基对（bp）的不完美反向重复序列，一个27bp的完美反向重复序列和一个17bp的富含A / T的区域。 DNA复制不需要T抗原结合位点I，但是它的存在使复制效率在体外和体内均提高了几倍。尽管SP1结合位点和增强子对体外复制几乎没有影响，但任何一种元素的存在均显着提高了体内复制。因此，这些元件的生物学作用不限于刺激转录，而是可能更普遍。

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