慢生根瘤菌的nifH和nifDK操纵子的激活取决于启动子上游的DNA序列。
Activation of the Bradyrhizobium japonicum nifH and nifDK operons is dependent on promoter-upstream DNA sequences.
作者信息
Alvarez-Morales A, Betancourt-Alvarez M, Kaluza K, Hennecke H
出版信息
Nucleic Acids Res. 1986 May 27;14(10):4207-27. doi: 10.1093/nar/14.10.4207.
Abstract
Previous analysis of B. japonicum nifH'- and nifD'-'lacZ translational fusions showed that these promoters could be activated by the K. pneumoniae nifA plus the E. coli ntrA gene products. To study the functions of the DNA 5' to these promoters, plasmids carrying deletions in this region were constructed and analyzed in vivo in a heterologous system consisting of an E. coli (NtrA+) background with a plasmid that constitutively expresses the K. pneumoniae nifA gene. Activation of the B. japonicum promoters was completely dependent on sequences located between positions -165 and -100, relative to the start of transcription. Some of the nifD deletion-fusions were mobilized to the wild-type B. japonicum and the exconjugants tested in an ex planta micro-aerobic system, and also used to infect soybean seedlings. The time course of derepression was followed by assaying beta-galactosidase activity from samples withdrawn from the microaerobic cultures or from root-nodule extracts. The results conclusively show that in the homologous system the sequences upstream of the promoter are required to achieve wild-type activity.
摘要
先前对日本固氮菌nifH'-和nifD'-'lacZ翻译融合体的分析表明，这些启动子可被肺炎克雷伯菌nifA加上大肠杆菌ntrA基因产物激活。为了研究这些启动子5'端DNA的功能，构建了在该区域带有缺失的质粒，并在由表达肺炎克雷伯菌nifA基因的质粒和大肠杆菌（NtrA+）背景组成的异源系统中进行体内分析。日本固氮菌启动子的激活完全依赖于相对于转录起始位置-165至-100之间的序列。一些nifD缺失融合体被转移到野生型日本固氮菌中，对接合后体在植物外微需氧系统中进行测试，并用于感染大豆幼苗。通过检测从微需氧培养物或根瘤提取物中取出的样品的β-半乳糖苷酶活性来跟踪去阻遏的时间进程。结果确凿地表明，在同源系统中，启动子上游的序列是实现野生型活性所必需的。
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