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9 This type of plasmid would seem to be ideal, but because all of the 2-tzm plasmid functions are required to maintain a stable copy number, 9 the cells must also contain endogenous 2-/~m circle plasmid (otherwise the plasmid behaves as an ARS plasmid, usually with a lower copy number). The presence of both plasmids in the cell is likely to result in problems with copurification and a risk of recombination between the plasmids resulting in hybrid plasmids. 3 kb), 1° although this would result in a large minichromosome (see later).
139. IRL Press, Oxford, 1998. 32F. Thoma and M. Zatchej, Cell 55, 945 (1988). 50 ASSEMBLY OF NUCLEOSOMES, CHROMATIN, AND NUCLEI  C h r o m a t i n  Assembly in Xenopus Extracts By DAVID JOHN TREMETHICK Introduction The majority of D N A in eukaryotic cells is packaged by histones, and many nonhistone proteins of unknown function, to form a dynamic structure known as chromatin. Chromatin is a periodic structure made up of repeating, regularly spaced subunits, the nucleosomes. Elegant genetic experiments have clearly demonstrated that histones play a central role in transcriptional control) Histones, via protein-protein interactions or by playing an architectural role, can facilitate or inhibit the transcriptional activation process.
18 A. Toh-e and R. B. Wickner, J. Bact. 145, 1421 (1981). 19 E. Erhart and C. P. Hollenberg, J. Bact. 156, 625 (1983). 20 S. M. Baker, P. G. Okhema, and J. A. Jaehning, Mol. Cell. Biol. 4, 2062 (1984). zl j. D. Boeke, F. LaCroute and G. R. Fink, Mol. Gen. Genet. 197, 345 (1984). 7% (w/v) agarose gel containing ethidium bromide. 7 kb relative to linear DNA. 5. GAL-FLP). Pick a single colony. Transformation of Yeast Cells Transformation of yeast cells may be achieved by the lithium acetate method 22 or by electroporation.