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在部分嘌呤饥饿期间诱导的枯草芽孢杆菌基因组lacZ融合体的分离与鉴定。

Isolation and characterization of Bacillus subtilis genomic lacZ fusions induced during partial purine starvation.

作者信息

Saxild H H, Jensen C L, Hubrechts P, Hammer K

机构信息

Department of Microbiology, Technical University of Denmark, Lyngby.

出版信息

J Bacteriol. 1994 Jan;176(2):276-83. doi: 10.1128/jb.176.2.276-283.1994.

DOI:10.1128/jb.176.2.276-283.1994
PMID:8288519
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC205047/
Abstract

Random genomic Bacillus subtilis lacZ fusions were screened in order to identify the possible existence of regulons responding to the stimuli generated by partial purine starvation. A leaky pur mutation (purL8) was isolated and used to generate the partial purine starvation conditions in the host strain used for screening. On the basis of their induction during partial purine starvation, seven genomic lacZ fusions were isolated. None of the fusions map in loci previously reported to contain purine-regulated genes. One fusion maps very close to the citB locus and may very well be a citB fusion. The fusions were divided into two types on the basis of their response to complete starvation for either ATP or GTP or both components at the same time. Except for one, type 2 fusions were induced by specific starvation for ATP and by simultaneous starvation for ATP and GTP, but not by specific GTP starvation in a gua strain or by GTP starvation induced by the addition of decoyinine. Type 1 fusions were equally well induced by all three kinds of purine starvation including GTP starvation induced by decoyinine. Further subdivisions of the fusions were obtained on the basis of their responses to the spo0A gene product. A total of five fusions showed that spo0A affected expression. One class was unable to induce lacZ expression in the absence of the spo0A gene product, whereas the other class had increased lacZ expression during partial purine starvation in a spo0A background.

摘要

为了确定是否存在对部分嘌呤饥饿所产生刺激作出反应的调节子,对随机基因组枯草芽孢杆菌lacZ融合体进行了筛选。分离出一个渗漏型嘌呤突变体(purL8),并用于在用于筛选的宿主菌株中产生部分嘌呤饥饿条件。基于它们在部分嘌呤饥饿期间的诱导情况,分离出了7个基因组lacZ融合体。这些融合体均不在先前报道的含有嘌呤调节基因的位点上定位。有一个融合体定位非常靠近citB位点,很可能是一个citB融合体。根据它们对ATP或GTP完全饥饿或同时对两者完全饥饿的反应,将这些融合体分为两种类型。除了一个之外,2型融合体在ATP特异性饥饿以及ATP和GTP同时饥饿时被诱导,但在gua菌株中GTP特异性饥饿或添加脱氧精胍菌素诱导的GTP饥饿时不被诱导。1型融合体在所有三种嘌呤饥饿情况下,包括脱氧精胍菌素诱导的GTP饥饿,均能被同样良好地诱导。根据它们对spo0A基因产物的反应,对融合体进行了进一步细分。总共有5个融合体表明spo0A影响表达。一类在没有spo0A基因产物时无法诱导lacZ表达,而另一类在spo0A背景下的部分嘌呤饥饿期间lacZ表达增加。

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