title: Rapid purification of HU protein fromHalobacillus karajensis.
abstract: The histone-like protein HU is the most-abundant DNA-binding protein in bacteria. The HU protein non-specifically binds and bends DNA as a hetero- or homodimer, and can participate in DNA supercoiling and DNA condensation. It also takes part in DNA functions such as replication, recombination, and repair. HU does not recognize any specific sequences but shows a certain degree of specificity to cruciform DNA and repair intermediates such as nick, gap, bulge, etc. To understand the features of HU binding to DNA and repair intermediates, a fast and easy HU protein purification method is required. Here we report a two-step purification procedure of HU fromHalobacillus karajensis(the gram positive and moderately halophilic bacteria isolated from Karaj surface soil). The method of HU purification allows obtaining a pure non-tagged protein. Salting out and ion exchange chromatography were applied for purification, and the purified protein was identified by immunoblotting. Results showed that the molecular weight of the purified protein was approximately 11 kDa which is immunologically similar to theBacillus subtilisHU protein (HBsu).
authors.affiliation: Department of Biology, Faculty of sciences, Alzahra University, Tehran, Iran.
authors.forename: Parinaz
authors.initials: P
authors.lastname: Ghadam
authors.affiliation: Department of Biology, Faculty of sciences, Alzahra University, Tehran, Iran.
authors.forename: Rana
authors.initials: R
authors.lastname: Samadi
keywords: DNA binding proteins; HBsu; HU; Halobacillus karajensis; Histone-like protein