Nematode chromosomal proteins--I. Isolation of chromatin and preliminary characterization of the chromosomal proteins of Caenorhabditis elegans.
1. A procedure is described which gives clean chromatin preparations from the free-living nematode Caenorhabditis elegans. It involves homogenization using glass beads, collection of the precipitate from a low speed centrifugation, removal of cell membranes with Triton X-100, several washes with 0.14 M NaCl, sucrose density gradient centrifugation, a cycle of extraction and reprecipitation using dilute Tris buffer and 0.14 M NaCl respectively, and final extraction of the purified deoxyribonucleoprotein in 10 mM Tris-HCl (pH 8). 2. Acidic urea gel electrophoresis of the histones from C. elegans yielded 4 main groups which were preliminary identified as H1, H2a (+ H3?), H2b, H4 and moved on the gels in that order of increasing mobility. the coincidence of histone H3 with H2a was putative, but its presence was firmly suggested by the generation of a dimeric form in oxidizing conditions. 3. By SDS-Tris-glycine gel electrophoresis of the non-histone chromosomal proteins of C. elegans, about 18 proteins were distinguished with molecular weights ranging from 15,000 to 100,000 daltons.
D000818:Animals; D002107:Caenorhabditis / Q000032:analysis* / Q000235:genetics; D002843:Chromatin / Q000302:isolation & purification*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000302:isolation & purification*; D012313:RNA / Q000032:analysis
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D012313:RNA; D004247:DNA