University of Illinois Urbana-Champaign

Construction and Characterization of Deletion Mutations in Domain C of ARSl

Woontner, Michael Roger

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Description

Title
Construction and Characterization of Deletion Mutations in Domain C of ARSl
Author(s)
Woontner, Michael Roger
Issue Date
1987
Doctoral Committee Chair(s)
Scott, John F.,
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 Sequences, or ARS elements, promote high-frequency transformation and extrachromosomal maintenance of plasmids in Saccharomyces cerevisiae, properties expected of DNA replication origins. A series of overlapping deletions in one flanking region (Domain C) of the ARSl element was constructed, and the effect of the deletions on the maintenance of various multicopy and single-copy plasmids examined. Analysis of the stabilities and copy numbers of multicopy plasmids indicated that while the core consensus element is absolutely required for extrachromosomal maintenance, the absence of Domain C has little effect. The loss rates of centromere (single-copy) plasmids increased slightly as the deletions approached the core consensus, suggesting that a block of sequence between 225 and 255 nucleotides from the consensus contains an element important to the maximal function of ARSl. These results also suggested that ARSl plays a role in replication, but has no effect on the segregation of centromere plasmids.
  • Comparison of multicopy plasmids of different sizes, each with an intact ARSl element, indicated a destabilizing effect of sequences derived from the E. coli cloning vector pBR322. A small amount of pBR322 (about a kilobase) was tolerated, and the bacterial ori was not responsible for the destabilizing effect.
Graduation Semester
1987
Type of Resource
text
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http://hdl.handle.net/2142/71188

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Dissertations and Theses - Microbiology