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

We have cloned a functional centromeric DNA sequence from Saccharomyces cerevisiae. Using the 2 mu chromosome-loss mapping technique and meiotic tetrad analysis, we have identified this DNA sequence as the centromere of chromosome V (CEN5). The CEN5 sequence has been localized on an 1,100-base-pair BamHI-BglII restriction fragment. Plasmids containing CEN5 and an autonomously replicating sequence are mitotically stable in S. cerevisiae and segregate in a Mendelian fashion during meiosis.

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Mutations at the Saccharomyces cerevisiae SUP4 tRNA Tyr Locus: Isolation, Genetic Fine-Structure Mapping, and Correlation with Physical Structure

Molecular and Cellular Biology ◽

10.1128/mcb.2.12.1501-1513.1982 ◽

1982 ◽

Vol 2 (12) ◽

pp. 1501-1513

Author(s):

Janet Kurjan ◽

Benjamin D. Hall

Keyword(s):

Saccharomyces Cerevisiae ◽

Fine Structure ◽

Physical Structure ◽

Mapping Technique ◽

Structure Mapping ◽

Isolation And Characterization ◽

Qualitative Estimate ◽

Genetic Fine Structure ◽

Entire Gene

The SUP4 tRNA Tyr locus in Saccharomyces cerevisiae has been studied by the isolation and characterization of mutations at the SUP4 gene which result in the loss of suppressor function. Most of the mutations act as single-site mutations, whereas about a third of the mutations are deletions of the entire gene. Two meiotic fine-structure maps of the gene were made. The first mapping technique placed 10 mutations plus the sup4 + anticodon on a map by a measurement of levels of recombination between pairs of mutations. The second map utilized a more qualitative estimate of recombination frequency, allowing 69 mutations and the sup4 + anticodon to be mapped. The maps were compared with the physical structure of the gene for the 34 mutations whose nucleotide alteration has been determined by DNA sequencing (Koski et al., Cell 22 :415-425, 1980; Kurjan et al., Cell 20 :701-709, 1980). Both maps show a good correlation with the physical structure of the gene, even though certain properties of genetic fine-structure maps, such as marker effects and “map expansion,” were seen.

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Isolation and characterization of an autonomously replicating sequence from Ustilago maydis

Molecular and Cellular Biology ◽

10.1128/mcb.8.9.3703-3709.1988 ◽

1988 ◽

Vol 8 (9) ◽

pp. 3703-3709

Author(s):

T Tsukuda ◽

S Carleton ◽

S Fotheringham ◽

W K Holloman

Keyword(s):

Saccharomyces Cerevisiae ◽

Consensus Sequence ◽

Ustilago Maydis ◽

Small Fragment ◽

Direct Repeats ◽

Base Pairs ◽

Autonomously Replicating Sequence ◽

Isolation And Characterization ◽

Extrachromosomal Elements

DNA fragments that function as autonomously replicating sequences (ARSs) have been isolated from Ustilago maydis. When inserted into an integrative transforming vector, the fragments increased the frequency of U. maydis transformation several-thousandfold. ARS-containing plasmids were transmitted in U. maydis as extrachromosomal elements through replication. They were maintained at a level of about 25 copies per cell but were mitotically unstable. One ARS characterized in detail, which we called UARS1, was localized to a 1.7-kilobase fragment. UARS1 contained a cluster of active sequences. This element could be reduced further into three separate subfragments, each of which retained ARS activity. The smallest one was 383 base pairs (bp) long. Although not active itself in yeast, this small fragment contained seven 8-bp direct repeats, two contiguous 30-bp direct repeats, and five 11-bp units in both orientations with sequences similar but not identical to the consensus sequence found to be crucial for ARS activity in Saccharomyces cerevisiae.

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Isolation and characterization of an autonomously replicating sequence from Ustilago maydis.

Molecular and Cellular Biology ◽

10.1128/mcb.8.9.3703 ◽

1988 ◽

Vol 8 (9) ◽

pp. 3703-3709 ◽

Cited By ~ 183

Author(s):

T Tsukuda ◽

S Carleton ◽

S Fotheringham ◽

W K Holloman

Keyword(s):

Saccharomyces Cerevisiae ◽

Consensus Sequence ◽

Ustilago Maydis ◽

Small Fragment ◽

Direct Repeats ◽

Base Pairs ◽

Autonomously Replicating Sequence ◽

Isolation And Characterization ◽

Extrachromosomal Elements

DNA fragments that function as autonomously replicating sequences (ARSs) have been isolated from Ustilago maydis. When inserted into an integrative transforming vector, the fragments increased the frequency of U. maydis transformation several-thousandfold. ARS-containing plasmids were transmitted in U. maydis as extrachromosomal elements through replication. They were maintained at a level of about 25 copies per cell but were mitotically unstable. One ARS characterized in detail, which we called UARS1, was localized to a 1.7-kilobase fragment. UARS1 contained a cluster of active sequences. This element could be reduced further into three separate subfragments, each of which retained ARS activity. The smallest one was 383 base pairs (bp) long. Although not active itself in yeast, this small fragment contained seven 8-bp direct repeats, two contiguous 30-bp direct repeats, and five 11-bp units in both orientations with sequences similar but not identical to the consensus sequence found to be crucial for ARS activity in Saccharomyces cerevisiae.

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