Nucleotide sequence comparisons and functional analysis of yeast centromere DNAs

Isolation and characterization of the centromere from chromosome V (CEN5) of Saccharomyces cerevisiae.

“…The CEN5 element stabilizes ARS-containing plasmids mitotically and meiotically (Tables 2 and 3), and, unlike CEN3 (4), CEN5 contains no ARS activity. The mitotic and meiotic stabilities of CEN5-containing plasmids were comparable to those of other CEN-containing plasmids (4,9,10,21,25). The CEN5 element was localized within a 1.1-kb BarnHI-BglII restriction fragment (Fig.…”

“…Most ARS-containing plasmids .n S. cerevisiae are unstable mitotically (26,27). Cloning of a centromeric DNA sequence into an ARScontaining plasmid results in the stable transmission of the plasmid to progeny during mitosis and meiosis (4,9,10,21,25 Table 2. Restfiction enzyme sites shown are: R, EcoRl; P, PstI; Bg, BgII; H3, HindIII; S, SalI; Bm, BamHI; Xb, XbaI; Hp, HpaI.…”

“…Like the other S. cerei'isiae centromeres which have been cloned (4,9,10,21,25), CEN5 is a unique DNA sequence in the yeast genome. The CEN5 element stabilizes ARS-containing plasmids mitotically and meiotically (Tables 2 and 3), and, unlike CEN3 (4), CEN5 contains no ARS activity.…”

“…3 days. Approximately 104 to 105 cells were inoculated into 10 ml of YEPD medium supplemented with 20 ,ug of uracil per ml and were grown to saturation at 30°C. Samples of the culture were plated to obtain single colonies, grown at 30°C, and replica plated onto CM-ura.…”

“…Plasmids containing a centromeric sequence and an autonomously replicating sequence (ARS) behave as minichromosomes which are mitotically stable and segregate 2+ :20 during meiosis (4,9,10,21,25). The centromere maintains the copy number of the plasmid at approximately one in a haploid cell (4).…”