Mapping and molecular cloning of the phn (psiD) locus for phosphonate utilization in Escherichia coli.
Mapping and molecular cloning of the phn (psiD) locus for phosphonate utilization in Escherichia coli.
复制标题
大肠杆菌中磷酸盐利用的 phn (psiD) 位点的定位和分子克隆。
DOI：
10.1128/jb.172.3.1186-1196.1990
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发表时间：
1990
影响因子：
3.2
通讯作者：
Boline,JA
中科院分区：
文献类型：
--
作者：
Wanner,BL;Boline,JA
大肠杆菌phn（psiD）基因座编码用于膦酸盐（Pn）利用的基因，因为phn（psiD）突变消除了使用具有取代的C-2或未取代的烃基团的Pn作为唯一P源的能力，所述取代的C-2或未取代的烃基团分别如2-氨基乙基膦酸盐（AEPn）或甲基膦酸盐（MPn）。尽管E. coliK-12磷酸饥饿诱导（psi）phn（psiD）基因表现正常的磷酸盐（Pi）控制，Pn利用在E. coliK-12以及E.大肠杆菌参考（ECOR）收集，与K-12密切相关。对于这些细菌，激活突变附近的phn（psiD）基因是必要的生长上的Pn作为唯一的P源。大多数E.大肠杆菌菌株，包括E. coli B，天然为Phn+;大肠杆菌菌株是Phn-，并且缺失了phn DNA序列。利用mini-Mu体内克隆技术和E. coliK-12 deltaphn突变体。phn（EcoB）DNA与E. coliK-12基因文库（Y.小原湾Akiyama和K. Isono，Cell 50：495-508，1987），其包含93-min区域，从而表明phn（psiD）基因座本身被克隆，并在其图谱位置上验证了我们的遗传数据。隐藏的phn（EcoK）DNA具有在天然Phn+ phn（EcoB）序列中不存在的另外100个碱基对。然而，在独立的Phn+ phn（EcoK）突变体中没有检测到总的结构变化，这些突变体在phn基因座附近具有激活突变。
The Escherichia coli phn (psiD) locus encodes genes for phosphonate (Pn) utilization, for phn (psiD) mutations abolish the ability to use as a sole P source a Pn with a substituted C-2 or unsubstituted hydrocarbon group such as 2-aminoethylphosphonate (AEPn) or methylphosphonate (MPn), respectively. Even though the E. coli K-12 phosphate starvation-inducible (psi) phn (psiD) gene(s) shows normal phosphate (Pi) control, Pn utilization is cryptic in E. coli K-12, as well as in several members of the E. coli reference (ECOR) collection which are closely related to K-12. For these bacteria, an activating mutation near the phn (psiD) gene is necessary for growth on a Pn as the sole P source. Most E. coli strains, including E. coli B, are naturally Phn+; a few E. coli strains are Phn- and are deleted for phn DNA sequences. The Phn+ phn(EcoB) DNA was molecularly cloned by using the mini-Mu in vivo cloning procedure and complementation of an E. coli K-12 delta phn mutant. The phn(EcoB) DNA hybridized to overlapping lambda clones in the E. coli K-12 gene library (Y. Kohara, K. Akiyama, and K. Isono, Cell 50:495-508, 1987) which contain the 93-min region, thus showing that the phn (psiD) locus was itself cloned and verifying our genetic data on its map location. The cryptic phn(EcoK) DNA has an additional 100 base pairs that is absent in the naturally Phn+ phn(EcoB) sequence. However, no gross structural change was detected in independent Phn+ phn(EcoK) mutants that have activating mutations near the phn locus.