Genetic Approaches to Study the Interplay Between Transcription and Nucleoid-Associated Proteins in Escherichia coli

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.