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Ludwigsen J, Klinker H, Mueller-Planitz F (2013) No need for a power stroke in ISWI-mediated nucleosome sliding. Mueller-Planitz F et al (2013) The ATPase domain of ISWI is an autonomous nucleosome remodeling machine. Maier VK, Becker PB (2012) A defined in vitro system to study ATP-dependent remodeling of short chromatin fibers. Maier VK et al (2008) ACF catalyses chromatosome movements in chromatin fibres. Nucleic Acids Res 34(10):2887–2905Įberharter A, Langst G, Becker PB (2004) A nucleosome sliding assay for chromatin remodeling factors. FEBS J 278(19):3579–3595įlaus A et al (2006) Identification of multiple distinct Snf2 subfamilies with conserved structural motifs. Annu Rev Biochem 78:273–304įlaus A, Owen-Hughes T (2011) Mechanisms for ATP-dependent chromatin remodelling: the means to the end. Cell Res 21(3):396–420Ĭlapier CR, Cairns BR (2009) The biology of chromatin remodeling complexes. Hargreaves DC, Crabtree GR (2011) ATP-dependent chromatin remodeling: genetics, genomics and mechanisms. Both assays exploit changes to the accessibility of DNA to restriction enzymes during the remodeling reaction. The first assay senses various structural changes to a specific nucleosome within the nucleosomal array whereas the second assay is specific toward detecting repositioning of nucleosomes within the array. Here, we describe how regularly spaced nucleosome arrays can be reconstituted in vitro and how these arrays can be used to dissect remodeling in the test tube. In vivo, however, nucleosomes form extended arrays with regular spacing. Remodeling has traditionally been analyzed using mononucleosomes as a model substrate. They deposit or evict nucleosomes or reposition them along DNA in a process termed nucleosome sliding. Isoschizomers offer flexibility of enzyme choice in situations where multiple enzymes are needed but do not have compatible buffers or reaction conditions.ATP-dependent nucleosome remodeling factors sculpt the nucleosomal landscape of eukaryotic chromatin. Isoschizomers have the same recognition site but may not have the same cut site. Two (or more) restriction enzymes may recognize the same sequence. Enzyme inactivation after digestion can also help prevent star activity. Star activity is when an enzyme begins to be less specific about the sites it cleaves. Using the correct reaction conditions and incubating for the proper length of time are important to prevent star activity in some enzymes. Restriction enzymes are typically sold with buffers in which they have optimal activity. These conditions include length and temperature of incubation. Restriction maps are invaluable for cloning, DNA typing, and any other experiment making use of restriction enzymes.Įach restriction enzyme has reaction conditions in which it works the best. A map of a DNA sequence showing the restriction sites present in that sequence is referred to as a restriction map. Restriction enzymes cleave double-stranded DNA within or adjacent to these specific sequences. Restriction enzymes (Restriction Endonucleases) recognize specific, short DNA sequences called recognition sequences, or restriction sites.

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