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T7 RNA聚合酶在T7和T3噬菌体染色体上读取的主要转录单元的初步图谱。

A preliminary map of the major transcription units read by T7 RNA polymerase on the T7 and T3 bacteriophage chromosomes.

作者信息

Golomb M, Chamberlin M

出版信息

Proc Natl Acad Sci U S A. 1974 Mar;71(3):760-4. doi: 10.1073/pnas.71.3.760.

Abstract

Transcription of T7 DNA by T7 RNA polymerase in vitro gives rise to six major size classes of RNAs comprising seven major T7 RNA species. These RNAs are all read from the r-strand of T7 DNA and are not derived from the early (leftmost on the conventional genetic map) region of the molecule. When artifically shortened T7 DNA templates are transcribed, four (I, II, IIIb, and VI) of the seven species are found to be truncated or deleted. This indicates that all are terminated near the right end of the T7 DNA molecule, probably at a common termination site near 98.5%. (Map positions are all given in terms of percentage of total length measured from the left end of the molecule.) Since the approximate lengths of the transcripts are known, the promotor sites for T7 RNA species I, II, IIIb, and VI are tentatively mapped at 56, 64, 83, and 97% on the T7 chromosome. Only a single major T3 RNA is transcribed by T7 RNA polymerase; analysis of transcripts directed by shortened T3 DNA templates indicates it is analogous to T7 RNA species IIIb. Hence the promotor and terminator sites for T3 species IIIb are tentatively mapped at 83 and 98.5%, respectively, on the T3 chromosome. The major transcripts read by T7 RNA polymerase from T3-T7 hybrid phage DNAs vary, depending on which regions of the T7 chromosome are present. This provides an alternative method of mapping the strong T7 promotor sites on the T7 chromosome.

摘要

T7 RNA聚合酶在体外对T7 DNA进行转录会产生六个主要大小类别的RNA,包括七种主要的T7 RNA种类。这些RNA均从T7 DNA的r链读取,并非源自该分子的早期(在传统遗传图谱上最左侧)区域。当对人工缩短的T7 DNA模板进行转录时,发现七种RNA中的四种(I、II、IIIb和VI)被截断或缺失。这表明所有这些RNA都在T7 DNA分子右端附近终止,可能在接近98.5%的一个共同终止位点。(图谱位置均以从分子左端测量的总长度的百分比表示。)由于已知转录本的大致长度,T7 RNA种类I、II、IIIb和VI的启动子位点暂定于T7染色体上的56%、64%、83%和97%处。T7 RNA聚合酶仅转录一种主要的T3 RNA;对缩短的T3 DNA模板指导的转录本分析表明,它类似于T7 RNA种类IIIb。因此,T3种类IIIb的启动子和终止子位点暂定于T3染色体上的83%和98.5%处。T7 RNA聚合酶从T3 - T7杂交噬菌体DNA读取的主要转录本有所不同,这取决于T7染色体的哪些区域存在。这提供了一种在T7染色体上定位强T7启动子位点的替代方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ccd6/388093/b73eee283b9d/pnas00056-0175-a.jpg

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