Abstract：
Bacterial nucleoid-associated proteins are important in nucleoid-structuring, homeostasis of DNA supercoiling, and in regulation of transcription. Vice versa, transcription influences DNA supercoiling and possibly DNA-binding of nucleoid-associated proteins. Here, I describe genetic tools to study the interplay between transcription and nucleoid-associated proteins such as H-NS in Escherichia coli. These standard methods include construction of genomic promoter reporter gene fusions to study regulation of promoters, genome insertion of promoter cassettes to drive expression of a gene of interest, and construction of isogenic hns mutants and precautions when doing so.
摘要：
细菌类核苷相关蛋白在核苷结构中很重要，DNA超螺旋的稳态，以及转录的调节。反之亦然,转录会影响DNA超螺旋作用，并可能影响类核苷酸相关蛋白的DNA结合。这里,我描述了研究大肠杆菌中转录和类核相关蛋白如H-NS之间相互作用的遗传工具。这些标准方法包括构建基因组启动子报告基因融合体以研究启动子的调控，启动子盒的基因组插入，以驱动感兴趣的基因的表达，以及等基因hns突变体的构建和预防措施。
