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Title:Construction and Characterization of Deletion Mutations in Domain C of ARSl
Author(s):Woontner, Michael Roger
Doctoral Committee Chair(s):Scott, John F.,
Department / Program:Microbiology
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(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.
Issue Date:1987
Description:115 p.
Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1987.
Other Identifier(s):(UMI)AAI8803241
Date Available in IDEALS:2014-12-16
Date Deposited:1987

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