Davey M E, de Bruijn F J
NSF Center for Microbial Ecology, MSU-DOE Plant Research Laboratory, and Department of Microbiology, Michigan State University, East Lansing, Michigan 48824, USA.
Appl Environ Microbiol. 2000 Dec;66(12):5353-9. doi: 10.1128/AEM.66.12.5353-5359.2000.
A nutrient deprivation-induced locus in Sinorhizobium meliloti strain 1021 was identified by use of a Tn5-luxAB reporter gene transposon. The tagged locus is comprised of two open reading frames (ORFs) designated ndiA and ndiB for nutrient deprivation-induced genes A and B. Comparison of the deduced amino acid sequences of both ndiA and ndiB to the protein databases failed to reveal similarity to any known genes. The expression of the ndi locus was found to be induced by carbon and nitrogen deprivation, osmotic stress, and oxygen limitation and during entry into stationary phase. To identify regulatory components involved in the control of ndi gene expression, a second round of mutagenesis was performed on the primary ndiB::Tn5-luxAB-tagged strain (C22) with transposon Tn1721. A double-mutant strain was obtained that lacked ndi locus transcriptional activity under all of the inducing conditions tested. The Tn1721-tagged gene showed a high degree of similarity to tryptophan-rich sensory protein TspO from Rhodobacter sphaeroides, as well as to mitochondrial benzodiazepine receptor pK18 from mammals. Induction of the ndi::Tn5-luxAB reporter gene fusion was restored under all inducing conditions by introducing the tspO coding region, from either S. meliloti or R. sphaeroides, in trans. Furthermore, it was found that, in addition to tspO, fixL, which encodes the sensor protein of an oxygen-sensing two-component system, is required for full expression of the ndi locus, but only under low oxygen tension.
利用Tn5-luxAB报告基因转座子,在苜蓿中华根瘤菌1021菌株中鉴定出一个营养剥夺诱导位点。该标记位点由两个开放阅读框(ORF)组成,分别命名为ndiA和ndiB,代表营养剥夺诱导基因A和B。将ndiA和ndiB推导的氨基酸序列与蛋白质数据库进行比较,未发现与任何已知基因有相似性。发现ndi位点的表达受碳氮剥夺、渗透胁迫、氧限制诱导,且在进入稳定期时也会被诱导。为了鉴定参与ndi基因表达调控的成分,用转座子Tn1721对初级ndiB::Tn5-luxAB标记菌株(C22)进行第二轮诱变。获得了一个双突变菌株,在所有测试的诱导条件下均缺乏ndi位点的转录活性。Tn1721标记的基因与球形红杆菌富含色氨酸的传感蛋白TspO以及哺乳动物的线粒体苯二氮䓬受体pK18高度相似。通过反式导入来自苜蓿中华根瘤菌或球形红杆菌的tspO编码区,在所有诱导条件下均可恢复ndi::Tn5-luxAB报告基因融合的诱导。此外,还发现,除了tspO外,编码氧传感双组分系统传感蛋白的fixL也是ndi位点充分表达所必需的,但仅在低氧张力下如此。