A tandem insertion vector for large-scale preparation of nucleosomal DNA

We developed a novel nucleosome DNA template vector, pWMD01, which is optimized for the large-scale preparation of nucleosomal DNA. By using restricted digestion by Sapl or Earl within its multicloning site, multiple half-nucleosome DNA units can be introduced unidirectionally into the vector at each subcloning step. Through this method, we constructed a plasmid that has 18 tandem repeats of a half-nucleosome 90-bp DNA unit containing c-Myb-binding sites in two subcloning cycles. This method enables the rapid, large-scale preparation of nucleosomal DNA with crystallization-grade quality. (C) 2012 Elsevier Inc. All rights reserved.