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噬菌体82基因Q和Q蛋白。序列、过量表达及其作为体外转录抗终止子的活性。

Bacteriophage 82 gene Q and Q protein. Sequence, overproduction, and activity as a transcription antiterminator in vitro.

作者信息

Goliger J A, Roberts J W

出版信息

J Biol Chem. 1987 Aug 25;262(24):11721-5.

PMID:3624233
Abstract

Phage 82 gene Q encodes a phage-specific positive regulator of late gene expression, thought, by analogy to the corresponding gene of phage lambda, to be a transcription antiterminator. We have cloned and sequenced the phage 82 gene Q and have overproduced and purified the 82 Q protein. We also have identified and sequenced DNA containing the phage 82 late gene promoter and terminator. We show that purified 82 Q protein is active and specific for DNA containing the 82 late gene promoter in a well defined in vitro transcription reaction: RNA polymerase initiating at the phage 82 late gene promoter and modified by 82 Q protein reads through a downstream transcriptional terminator. We used T1 RNase mapping to confirm that the putative readthrough RNA made in the presence of 82 Q protein is in fact an elongation product of the shorter RNA.

摘要

噬菌体82基因Q编码一种噬菌体特异性的晚期基因表达正调控因子,根据与噬菌体λ相应基因的类比,被认为是一种转录抗终止因子。我们克隆并测序了噬菌体82基因Q,过量表达并纯化了82 Q蛋白。我们还鉴定并测序了包含噬菌体82晚期基因启动子和终止子的DNA。我们表明,在明确的体外转录反应中,纯化的82 Q蛋白对含有82晚期基因启动子的DNA具有活性且具有特异性:在噬菌体82晚期基因启动子处起始并被82 Q蛋白修饰的RNA聚合酶能够通读下游的转录终止子。我们使用T1核糖核酸酶图谱分析来确认在82 Q蛋白存在下产生的推定通读RNA实际上是较短RNA的延伸产物。

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