Affinity Purification of Specific Chromatin Segments from Chromosomal Loci in Yeast

Abstract

Single-copy gene and promoter regions have been excised from yeast chromosomes and have been purified as chromatin by conventional and affinity methods. Promoter regions isolated in transcriptionally repressed and activated states maintain their characteristic chromatin structures. Gel filtration analysis establishes the uniformity of the transcriptionally activated state. Activator proteins interact in the manner anticipated from previous studies in vivo. This work opens the way to the direct study of specific gene regions of eukaryotic chromosomes in diverse functional and structural states.

ACKNOWLEDGMENTS

Plasmids pRS415-RecR and pABX22 were a kind gift from M. Gartenberg. Plasmid pPHO2-His was a kind gift from D. Stillman.

This research was supported by NIH grant GM36659 to R.D.K.

J.G. and H.B. contributed equally to this work.