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大肠杆菌噬菌体T5基因组的结构与功能。“有切口的”和“无切口的”T5+ DNA的体外转录。

Structure and function of the genome of coliphage T5. Transcription in vitro of the "nicked" and "nick-free" T5+ DNA.

作者信息

Knopf K, Bujard H

出版信息

Eur J Biochem. 1975 May 6;53(2):371-85. doi: 10.1111/j.1432-1033.1975.tb04077.x.

Abstract

Purified T5+ DNA in nicked form and after repair of the specific single-strand interruptions with DNA ligase was used in transcription studies using Escherichia coli RNA polymerase (holoenzyme) in the presence and absence of E. coli termination protein rho. The transcriptional products were analyzed with respect to their size distribution and the sequences transcribed from the different templates. The results indicate that the single-strand breaks in the DNA of bacteriophage T5, though in genetically defined positions, do not have any specific effect on transcription in vitro. Furthermore, the E. coli rho protein, although it depresses net RNA synthesis and reduces the average molecular weight of the transcripts, seems to act in a non-specific way in this system.

摘要

以带切口形式存在的纯化T5 + DNA,以及用DNA连接酶修复特定单链中断后的DNA,用于转录研究。该研究使用大肠杆菌RNA聚合酶(全酶),分别在有和没有大肠杆菌终止蛋白rho的情况下进行。针对转录产物的大小分布以及从不同模板转录的序列进行了分析。结果表明,噬菌体T5 DNA中的单链断裂,尽管处于基因定义的位置,但对体外转录没有任何特定影响。此外,大肠杆菌rho蛋白虽然会抑制净RNA合成并降低转录本的平均分子量,但在该系统中似乎以非特异性方式起作用。

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