Structure and Function of the Yeast Chromosomal Replicator Ars1 (Plasmid)

Strich, Randy Stephen

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Description

Title

Structure and Function of the Yeast Chromosomal Replicator Ars1 (Plasmid)

Author(s)

Strich, Randy Stephen

Issue Date

1986

Department of Study

Microbiology

Discipline

Microbiology

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

Biology, Molecular

Abstract

Autonomously Replicating Sequence 1 (ARS1) is a cis acting DNA sequence that allows the extra-chromosomal maintenance of plasmids in the yeast Saccharomyces cerevisiae. In order to define the functional domains of ARS1 and determine its role in plasmid maintenance, I have performed in vitro mutagenesis in the region containing ARS1 and measured their effects on Ars activity using the parameters of high-frequency transformation, colony morphology, copy number, and stability in both high- and low-copy plasmids. The mutagenesis was in the form of insertions (10-4200 base pairs) and deletions. My results indicate that the consensus sequence found in all ARS elements is essential for Ars function and that one flanking region but not the other is important for maximizing Ars activity. The effects of the mutations on plasmid maintenance include altered colony morphology due to severe plasmid loss, reduced copy number, a reduction in the percent of plasmid-bearing cells using high-copy plasmids, and an increased rate of 1:0 (simple loss) using low-copy centromere containing plasmids. These results indicate that the role ARS1 plays in plasmid maintenance is concerned with the replication and/or localization of the plasmid and not its segregation.

Graduation Semester

1986

Type of Resource

text

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http://hdl.handle.net/2142/71180

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