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[噬菌体T7 RNA聚合酶:基因克隆及其结构]

[Phage T7 RNA-polymerase: gene cloning and its structure].

作者信息

Grachev M A, Pletnev A G

出版信息

Bioorg Khim. 1984 Jun;10(6):824-43.

PMID:6093820
Abstract

Gene 1.0 to T7 phage coding for a phage-specific RNA polymerase has been cloned in a plasmid vector. One of the clones obtained, viz. E. coli K12 HB101 (pSK-T7-1.0a), produced an active T7 RNA polymerase as revealed by the fact that its extract stimulated RNA synthesis on the T7 DNA template in the presence of rifampicin. Analysis of gene 1.0 fragments isolated from pSK-T7-1.0a plasmid made it possible to refine the sequence of this gene which has been recently published by the authors of the present paper and, independently, by other workers. Another plasmid, pSK-T7-1.0b, did not differ from pSK-T7-1.0a in the restriction map. However, it failed to induce production of a rifampicin-insensitive RNA polymerase. Sequencing revealed a deletion of one T residue in gene 1.0 present in this plasmid.

摘要

编码噬菌体特异性RNA聚合酶的T7噬菌体的基因1.0已被克隆到质粒载体中。获得的其中一个克隆,即大肠杆菌K12 HB101(pSK-T7-1.0a),产生了一种活性T7 RNA聚合酶,这一事实表明,在利福平存在的情况下,其提取物刺激了T7 DNA模板上的RNA合成。对从pSK-T7-1.0a质粒分离的基因1.0片段的分析使得能够完善该基因的序列,该序列最近由本文作者以及其他研究人员独立发表。另一个质粒pSK-T7-1.0b在限制性图谱上与pSK-T7-1.0a没有差异。然而,它未能诱导产生对利福平不敏感的RNA聚合酶。测序显示该质粒中存在的基因1.0中有一个T残基缺失。

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