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Creation of an aconitase overexpression strain of Saccharomyces cerevisiae for lifespan analysis

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Date Issued:
2012
Summary:
In my thesis work, I attempted to construct a plasmid that would allow stable integration of genes into the Saccharomyces cerevisiae yeast genome under the control of the repressible TetO promoter. The yeast ACO1 gene was cloned under the control of the TetO operator and the tTA transactivator. This construct was inserted into yeast cells in order to observe the effects of aconitase overexpression on aging. Unfortunately, the transformed cells appeared incapable of aconitase expression as determined by glutamic acid auxptrophy, a phenotype of aconitase mutants. We have sequenced the pIT1ACO1 plasmid and have found many abnormalities in the promoter region. If the plasmid can be made to function as intended, the resulting yeast strain can be used in the future to determine if aconitase plays an important role in cellular aging.
Title: Creation of an aconitase overexpression strain of Saccharomyces cerevisiae for lifespan analysis.
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Name(s): Nunes, Steve.
Harriet L. Wilkes Honors College
Type of Resource: text
Genre: Thesis
Issuance: multipart monograph
Date Issued: 2012
Publisher: Florida Atlantic University
Physical Form: electronic
electronic resource
Extent: v, 20 p. : ill. (some col.)
Language(s): English
Summary: In my thesis work, I attempted to construct a plasmid that would allow stable integration of genes into the Saccharomyces cerevisiae yeast genome under the control of the repressible TetO promoter. The yeast ACO1 gene was cloned under the control of the TetO operator and the tTA transactivator. This construct was inserted into yeast cells in order to observe the effects of aconitase overexpression on aging. Unfortunately, the transformed cells appeared incapable of aconitase expression as determined by glutamic acid auxptrophy, a phenotype of aconitase mutants. We have sequenced the pIT1ACO1 plasmid and have found many abnormalities in the promoter region. If the plasmid can be made to function as intended, the resulting yeast strain can be used in the future to determine if aconitase plays an important role in cellular aging.
Identifier: 819413214 (oclc), 3359306 (digitool), FADT3359306 (IID), fau:1434 (fedora)
Note(s): by Steve Nunes.
Thesis (B.A.)--Florida Atlantic University, Honors College, 2012.
Includes bibliography.
Electronic reproduction. Boca Raton, Fla., 2012. Mode of access: World Wide Web.
Subject(s): Yeast fungi -- Research -- Methodology
Microbial genetics
Genetic engineering
Aging -- Molecular aspects
Cell metabolism
Mutation (Biology)
Held by: FBoU FAUER
Persistent Link to This Record: http://purl.flvc.org/FAU/3359306
Use and Reproduction: Copyright © is held by the author, with permission granted to Florida Atlantic University to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Host Institution: FAU

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