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Host-specific activation of transcription by tandem repeats from simian virus 40 and Moloney murine sarcoma virus.

机译:串联重复的宿主特异性激活猿猴病毒40和莫洛尼鼠肉瘤病毒。

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摘要

The simian virus (SV40) 72-base pair (bp) tandem repeated sequences have recently been shown to function as activators or enhancers of early viral transcription. A recombinant viral genome was recently constructed by inserting 72-bp tandem repeats from the Moloney murine sarcoma virus (MSV) in place of the 72-bp repeats of SV40. Although this genome replicates in monkey kidney cells, its rate of large tumor antigen expression and replication is considerably slower than that of wild-type SV40. In mouse cells, however, equivalent levels of large tumor antigen appear to be expressed from both wild-type and recombinant genomes, suggesting a relationship between the level of enhancer activity and the host cell. To confirm this observation, we have applied a sensitive quantitative assay for gene expression based on the conversion of chloramphenicol to its acetylated forms. The gene encoding the enzymatic function chloramphenicol acetyltransferase was inserted into two vectors in which the enhancer sequences from SV40 or MSV were placed adjacent to the early SV40 promoter. The SV40 tandem repeats appear to activate gene expression to significantly higher levels in monkey kidney cells, but the MSV repeats are more active in two lines of mouse cells. These findings suggest that the tandem repeat elements may interact with host-specific molecules and, furthermore, may constitute one of the elements determining the host range of these eukaryotic viruses.
机译:猿猴病毒(SV40)72个碱基对(bp)的串联重复序列最近已显示出可作为早期病毒转录的激活剂或增强剂。最近,通过插入莫洛尼鼠肉瘤病毒(MSV)的72 bp串联重复序列代替SV40的72 bp重复序列,构建了重组病毒基因组。尽管该基因组在猴肾细胞中复制,但是其大肿瘤抗原表达和复制的速度比野生型SV40慢得多。然而,在小鼠细胞中,野生型和重组基因组均表达了同等水平的大肿瘤抗原,这表明增强子活性水平与宿主细胞之间存在关系。为了证实这一发现,我们基于氯霉素转化为其乙酰化形式,对基因表达进行了灵敏的定量分析。将编码酶功能氯霉素乙酰转移酶的基因插入两个载体,其中来自SV40或MSV的增强子序列与早期SV40启动子相邻放置。 SV40串联重复序列似乎可以在猴肾细胞中将基因表达激活至更高的水平,但是MSV重复序列在两行小鼠细胞中的活性更高。这些发现表明串联重复元件可以与宿主特异性分子相互作用,而且可以构成确定这些真核病毒宿主范围的元件之一。

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