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在枯草芽孢杆菌氮限制生长期间表达被激活的基因和基因产物的鉴定。

Identification of genes and gene products whose expression is activated during nitrogen-limited growth in Bacillus subtilis.

作者信息

Atkinson M R, Fisher S H

机构信息

Department of Microbiology, Boston University School of Medicine, Massachusetts 02118.

出版信息

J Bacteriol. 1991 Jan;173(1):23-7. doi: 10.1128/jb.173.1.23-27.1991.

Abstract

The levels of urease and asparaginase were elevated 25- and 20-fold, respectively, in extracts of Bacillus subtilis cells grown in medium containing nitrogen sources that are poor sources of ammonium (NH4+) compared with the levels seen in extracts of cells grown in medium containing nitrogen sources that are good sources of NH4+. To determine whether a collection of genes whose expression responds to nitrogen availability could be isolated, a library of Tn917-lacZ insertions was screened for nitrogen-regulated beta-galactosidase expression. Two fusion strains were identified. beta-Galactosidase expression was 26- and 4,000-fold higher, respectively, in the nrg-21::Tn917-lacZ and the nrg-29::Tn917-lacZ insertion strains during NH4(+)-restricted growth than during growth on nitrogen sources that are good sources of NH4+. PBS1 transduction analysis showed that the nrg-21::Tn917-lacZ insertion mapped between gutB and purB and that the nrg-29::Tn917-lacZ insertion mapped between degSU and spoIID. The repression of expression of these four gene products during growth on good sources of NH4+ required the wild-type glutamine synthetase protein but not the glutamine synthetase regulatory protein, GlnR.

摘要

与在含有丰富铵源(NH4+)的氮源培养基中生长的细胞提取物相比,在含有铵源匮乏的氮源培养基中生长的枯草芽孢杆菌细胞提取物中,脲酶和天冬酰胺酶的水平分别升高了25倍和20倍。为了确定是否可以分离出一组其表达对氮可用性有反应的基因,对Tn917 - lacZ插入文库进行了氮调节β-半乳糖苷酶表达筛选。鉴定出两个融合菌株。在NH4(+)受限生长期间,nrg - 21::Tn917 - lacZ和nrg - 29::Tn917 - lacZ插入菌株中的β-半乳糖苷酶表达分别比在含有丰富NH4+的氮源上生长时高26倍和4000倍。PBS1转导分析表明,nrg - 21::Tn917 - lacZ插入位于gutB和purB之间,nrg - 29::Tn917 - lacZ插入位于degSU和spoIID之间。在含有丰富NH4+的氮源上生长期间,这四种基因产物表达的抑制需要野生型谷氨酰胺合成酶蛋白,但不需要谷氨酰胺合成酶调节蛋白GlnR。

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