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T7和T3特异性RNA聚合酶:对从T3 DNA读取的体外转录本的表征与定位

T7- and T3-specific RNA polymerases: characterization and mapping of the in vitro transcripts read from T3 DNA.

作者信息

Golomb M, Chamberlin M J

出版信息

J Virol. 1977 Feb;21(2):743-52. doi: 10.1128/JVI.21.2.743-752.1977.

Abstract

The late genetic region of the T3 bacteriophage genome is transcribed by a phage-specified RNA polymerase, the product of T3 gene 1. In vitro, purified T3 RNA polymerase acting with T3 DNA template synthesizes six major RNA products, with molecular weights of 5.5 X 10(6), 4.2 X 10(6), 1.7 X 10(6), 0.87 X 10(6), 0.52 X 10(6), and 0.23 X 10(6). These are designated T3 RNA species I through VI, respectively. These RNAs are formed in equimolar amounts, with the exception of T3 species V, which is made in approximately twice this amount. T3 RNA species I, II, and VI have been mapped by the use of terminally deleted templates and are found to originate from promoters located at 56, 67, and 98% genome length, respectively, and to share a common terminator at approximately 100%. T3 species III, IV, and V must originate from the class III region of the T3 genome between 37.5 and 56%. Although the pattern of transcription by T3 RNA polymerase in vitro closely resembles that of the RNA polymerase from the related phage T7, neither polymerase is able to use major promoter sites for the other enzyme at an appreciable rate.

摘要

T3噬菌体基因组的晚期遗传区域由噬菌体特异的RNA聚合酶转录,该酶是T3基因1的产物。在体外,纯化的T3 RNA聚合酶与T3 DNA模板作用,合成六种主要的RNA产物,分子量分别为5.5×10⁶、4.2×10⁶、1.7×10⁶、0.87×10⁶、0.52×10⁶和0.23×10⁶。这些分别被指定为T3 RNA种类I至VI。除了T3种类V的产量约为其他种类的两倍外,这些RNA以等摩尔量形成。通过使用末端缺失的模板对T3 RNA种类I、II和VI进行了定位,发现它们分别起源于位于基因组长度56%、67%和98%处的启动子,并在大约100%处共享一个共同的终止子。T3种类III、IV和V必定起源于T3基因组37.5%至56%之间的III类区域。尽管T3 RNA聚合酶在体外的转录模式与相关噬菌体T7的RNA聚合酶非常相似,但这两种聚合酶都不能以可观的速率使用另一种酶的主要启动子位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9a5/353876/93f91d2fbec1/jvirol00206-0317-a.jpg

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