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一个依赖于cII的启动子位于噬菌体λ的Q基因内。

A cII-dependent promoter is located within the Q gene of bacteriophage lambda.

作者信息

Hoopes B C, McClure W R

出版信息

Proc Natl Acad Sci U S A. 1985 May;82(10):3134-8. doi: 10.1073/pnas.82.10.3134.

Abstract

We have found a cII-dependent promoter, PaQ, within the Q gene of bacteriophage lambda. Transcription experiments and abortive initiation assays performed in vitro showed that the promoter strength and the cII affinity of PaQ were comparable to the other cII-dependent lambda promoters, PE and PI. The location and leftward direction of PaQ suggests a possible role in the delay of lambda late-gene expression by cII protein, a phenomenon that has been called cII-dependent inhibition. We have constructed a promoter down mutation, paq-1, by changing a single base pair in the putative cII binding site of the promoter by oligonucleotide site-directed mutagenesis. The paq-1 mutant promoter required about 4-fold higher cII concentrations for maximal activation compared to the wild-type PaQ. We tested the hypothesis that PaQ is responsible in part for the delay of lambda late-gene expression by recombining the paq-1 mutation into a phage showing severe cII-dependent inhibition. We found that the paq-1 mutation relieved the cII-dependent growth defect of this phage. The paq-1 mutation (in combination with lambda cI857) resulted in a clear-plaque phenotype at the permissive temperature of 32 degrees C. The role of the PaQ-initiated antisense transcript in the control of lambda development is discussed.

摘要

我们在噬菌体λ的Q基因内发现了一个依赖于cII的启动子PaQ。体外进行的转录实验和流产起始分析表明,PaQ的启动子强度和对cII的亲和力与其他依赖于cII的λ启动子PE和PI相当。PaQ的位置和向左的方向表明它可能在cII蛋白延迟λ晚期基因表达中发挥作用,这一现象被称为cII依赖性抑制。我们通过寡核苷酸定点诱变改变启动子假定的cII结合位点中的一个碱基对,构建了一个启动子下调突变体paq-1。与野生型PaQ相比,paq-1突变体启动子需要约4倍高的cII浓度才能实现最大激活。我们通过将paq-1突变重组到一个表现出严重cII依赖性抑制的噬菌体中,来检验PaQ部分负责λ晚期基因表达延迟的假说。我们发现paq-1突变缓解了该噬菌体的cII依赖性生长缺陷。paq-1突变(与λcI857组合)在32℃的允许温度下产生了清晰噬菌斑表型。本文讨论了由PaQ起始的反义转录本在λ发育调控中的作用。

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