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Creation of an aconitase overexpression strain of Saccharomyces cerevisiae for lifespan analysis
Title
Creation of an aconitase overexpression strain of Saccharomyces cerevisiae for lifespan analysis.
Creator
Nunes, Steve., Harriet L. Wilkes Honors College
Abstract/Description
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...
Show moreIn 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.
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Date Issued
2012
PURL
http://purl.flvc.org/FAU/3359306
Subject Headings
Yeast fungi, Research, Methodology, Microbial genetics, Genetic engineering, Aging, Molecular aspects, Cell metabolism, Mutation (Biology)
Format
Document (PDF)
Devising a noncancerous model system to study multipolar spindle formation
Title
Devising a noncancerous model system to study multipolar spindle formation.
Creator
Nagarsheth, Nisha., Harriet L. Wilkes Honors College
Abstract/Description
Aneuploid tumor cells have characteristically unstable genomes which can be caused by mitotic defects such as multipolar spindles. Multipolarity relies upon the presence of extra centrosomes to form. However, some cells, both cancerous and noncancerous are able to avoid the formation of multipolar spindles through centrosomal clustering. Previous research has shown that there are a large number of genes whose activity contributes to the clustering activity, making analysis of individual...
Show moreAneuploid tumor cells have characteristically unstable genomes which can be caused by mitotic defects such as multipolar spindles. Multipolarity relies upon the presence of extra centrosomes to form. However, some cells, both cancerous and noncancerous are able to avoid the formation of multipolar spindles through centrosomal clustering. Previous research has shown that there are a large number of genes whose activity contributes to the clustering activity, making analysis of individual components of the process difficult. In order to better understand centrosomal clustering in cancer cells, we induced supernumerary centrosomes in a genomically normal cell line, RPE, to observe how the normal cells cope with extra centrosomes. Using colcemid to induce extra centrosomes in the RPE cell line, we observed an intact clustering mechanism in fixed cells. Further manipulation of the cells has allowed us to induce multipolarity in this cell line using various disrupters of cell-cycle checkpoint and dynein function.
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Date Issued
2010
PURL
http://purl.flvc.org/FAU/3335107
Subject Headings
Centrosomes, Research, Cancer, Genetic aspects, Cellular signal transduction, Cell division
Format
Document (PDF)
effect of mutated aconitase on yeast longevity
Title
The effect of mutated aconitase on yeast longevity.
Creator
Kwan, CJ., Harriet L. Wilkes Honors College
Abstract/Description
Aconitase is an important enzyme in the citric Acid Cycle, is needed for maintenance of mitochondrial DNA, is a key regulator of iron in the cell, and is very sensitive to oxidative stress. We have isolatd the yeast ACO1 gene, which codes for aconitase, and randomly mutated it to create a mutant library of cells each expressing a different version of ACO1. We will select for oxidative stress resistant aconitase in S. cerevisiae by subjecting strains to successive rounds of heat shock and...
Show moreAconitase is an important enzyme in the citric Acid Cycle, is needed for maintenance of mitochondrial DNA, is a key regulator of iron in the cell, and is very sensitive to oxidative stress. We have isolatd the yeast ACO1 gene, which codes for aconitase, and randomly mutated it to create a mutant library of cells each expressing a different version of ACO1. We will select for oxidative stress resistant aconitase in S. cerevisiae by subjecting strains to successive rounds of heat shock and competitive growth against other mutants. The "winner" of this competition will then be analyzed for which version of aconitase it is expressing, which may lead to increased longevity.
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Date Issued
2012
PURL
http://purl.flvc.org/FAU/3359310
Subject Headings
Yeast fungi, Research, Microbial genetics, Aging, Molecular aspects, Mutation (Biology), Cell metabolism
Format
Document (PDF)
House ants of Karachi, Pakistan
Title
House ants of Karachi, Pakistan.
Creator
Haji, Malik., Harriet L. Wilkes Honors College
Abstract/Description
Ants are ubiquitous and ecologically important, yet little is known of ants within Pakistan. I studied ants collected from houses and apartments in Karachi, Pakistan to determine which ants are common indoor ants. Ants were attracted by spilling syrups and sodas on the floor which were left overnight. The following morning the ants were collected using an aspirator. A total of eleven species were found. Of these eight were well-known pest ants with worldwide distributions: Paratrechina...
Show moreAnts are ubiquitous and ecologically important, yet little is known of ants within Pakistan. I studied ants collected from houses and apartments in Karachi, Pakistan to determine which ants are common indoor ants. Ants were attracted by spilling syrups and sodas on the floor which were left overnight. The following morning the ants were collected using an aspirator. A total of eleven species were found. Of these eight were well-known pest ants with worldwide distributions: Paratrechina longicornis, Tetramorium simillimum, Monomorium pharaonis, Monomorium subopacum, Monomorium destructor, Tapinoma melanocephalum, Pheidole megacephala, and Tetramorium bicarinatum. Three others, Camponotus sp., Pheidole sp., and Solenopsis sp., not identified to species were probably native species. Paratrechina longicornis and M. subopacum were the two most dominant species that were found in the highest abundance.
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Date Issued
2008
PURL
http://purl.flvc.org/FAU/77670
Subject Headings
Insects, Identification, Ants, Research, Self-organizing systems
Format
Document (PDF)
Iron and mitochondrial aging
Title
Iron and mitochondrial aging.
Creator
Paez, Rafael., Harriet L. Wilkes Honors College
Abstract/Description
Aging is a process characterized by accumulated oxidative damage to DNA, proteins, and lipids,which leads to the gradual degeneration of cellular activity. Mitochondria play a central role in aging because they produce both cellular energy and oxidative stress. As resultof accumulated oxidative damage, mitochondria function decays, which leads to a cellular energy deficit and compromises cellular function. Iron is an essential nutrient reequired by mitodhondria to function optimally. It has...
Show moreAging is a process characterized by accumulated oxidative damage to DNA, proteins, and lipids,which leads to the gradual degeneration of cellular activity. Mitochondria play a central role in aging because they produce both cellular energy and oxidative stress. As resultof accumulated oxidative damage, mitochondria function decays, which leads to a cellular energy deficit and compromises cellular function. Iron is an essential nutrient reequired by mitodhondria to function optimally. It has been proved that iron supplementation increases the lifespan of several yeast strains, including superoxide dismutase mutants. We are interested in finding where the iron is going and what it might be doing that is beneficial to the cell. We have used Saccharomyces cerevisiae as our molecular model of aging. Our results indicate that the extra iron is being transported into the mitochoindria.
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Date Issued
2012
PURL
http://purl.flvc.org/FAU/3359313
Subject Headings
Oxidation, Physiological, Aging, Molecular aspects, Mitochondrial DNA, Yeast fungi, Research, Methodology, Free radicals (Chemistry), Pathophysiology
Format
Document (PDF)