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Full-Text Articles in Cell Biology

A Novel Kinesin Adapter Directly Mediates Dendritic Mrna Localization During Synapse Development, Hao Wu May 2019

A Novel Kinesin Adapter Directly Mediates Dendritic Mrna Localization During Synapse Development, Hao Wu

All Dissertations, Theses, and Capstone Projects

Cytoskeleton based active transport with motor proteins is essential for mRNA localization and local protein translation in animal cells, yet how mRNA granules interact with motor proteins remains poorly understood. Using an unbiased screen for interaction between mRNA binding proteins (RBP) and motor proteins, we identified protein interacting with APP tail 1 (PAT1) as a potential direct adapter between the β-actin mRNA Zipcode-binding protein 1 (ZBP1) and Kinesin-1 motor complex.

Mouse PAT1 is similar to the Kinesin Light Chain (KLC) in amino acid sequence and binds directly to KLC. High-resolution images from structured illumination microscopy (SIM) indicates that synaptic stimulation ...


Genome-Scale Crispr Screens Identify Human Pluripotency-Specific Genes, Robert J. Ihry, Zinger Yang, Ajamete Kaykas Apr 2019

Genome-Scale Crispr Screens Identify Human Pluripotency-Specific Genes, Robert J. Ihry, Zinger Yang, Ajamete Kaykas

Open Access Articles

Human pluripotent stem cells (hPSCs) generate a variety of disease-relevant cells that can be used to improve the translation of preclinical research. Despite the potential of hPSCs, their use for genetic screening has been limited by technical challenges. We developed a scalable and renewable Cas9 and sgRNA-hPSC library in which loss-of-function mutations can be induced at will. Our inducible mutant hPSC library can be used for multiple genome-wide CRISPR screens in a variety of hPSC-induced cell types. As proof of concept, we performed three screens for regulators of properties fundamental to hPSCs: their ability to self-renew and/or survive (fitness ...


A Functional Unfolded Protein Response Is Required For Normal Vegetative Development, Yan Bao, Diane C. Bassham, Stephen H. Howell Apr 2019

A Functional Unfolded Protein Response Is Required For Normal Vegetative Development, Yan Bao, Diane C. Bassham, Stephen H. Howell

Genetics, Development and Cell Biology Publications

The unfolded protein response (UPR) is activated in plants in response to endoplasmic reticulum stress and plays an important role in mitigating stress damage. Multiple factors act in the UPR, including the membrane-associated transcription factor, BASIC LEUCINE ZIPPER 17 (bZIP17), and the membrane-associated RNA splicing factor, INOSITOL REQUIRING ENZYME1 (IRE1). We have analyzed an Arabidopsis (Arabidopsis thaliana) ire1a ire1b bzip17 triple mutant, with defects in stress signaling, and found that the mutant is also impaired in vegetative plant growth under conditions without externally applied stress. This raised the possibility that the UPR functions in plant development in the same manner ...


X-Inactivation And Epigenetics, Serena Weston Apr 2019

X-Inactivation And Epigenetics, Serena Weston

Thinking Matters Symposium

To prevent abnormal development caused by expressing both X chromosomes, female mammals inactivate one of their X chromosomes using an epigenetic process called dosage compensation. This literature review examines how X chromosome inactivation (XCI) occurs during the formation and development of an embryo. This condensation of DNA is marked by histone tail modifications, DNA methylation, and the arrival of structural proteins resulting in extraordinarily stable heterochromatin. X-inactivation is regulated in cis by the X-inactivation center (Xic) that contains the Xist gene and its antisense gene, Tsix. On one X chromosome, Xist RNA is expressed and coats the center of the ...


Unified Methods For Feature Selection In Large-Scale Genomic Studies With Censored Survival Outcomes, Lauren Spirko-Burns, Karthik Devarajan Mar 2019

Unified Methods For Feature Selection In Large-Scale Genomic Studies With Censored Survival Outcomes, Lauren Spirko-Burns, Karthik Devarajan

COBRA Preprint Series

One of the major goals in large-scale genomic studies is to identify genes with a prognostic impact on time-to-event outcomes which provide insight into the disease's process. With rapid developments in high-throughput genomic technologies in the past two decades, the scientific community is able to monitor the expression levels of tens of thousands of genes and proteins resulting in enormous data sets where the number of genomic features is far greater than the number of subjects. Methods based on univariate Cox regression are often used to select genomic features related to survival outcome; however, the Cox model assumes proportional ...


Investigating The Role Of Free Radicals In Huntington's Disease Using Drosophila Melanogaster, Jennifer Libov Mar 2019

Investigating The Role Of Free Radicals In Huntington's Disease Using Drosophila Melanogaster, Jennifer Libov

Honors Theses, University of Nebraska-Lincoln

During normal cell metabolism, reactive oxygen species (ROS) are produced as a byproduct of oxidative phosphorylation. ROS are utilized in the cell as a signaling molecule and can be maintained at healthy levels by cellular antioxidants. However, when the cell experiences oxidative stress due to environmental or genetic conditions, levels of ROS can exceed healthy levels and inhibit necessary life functions by damaging biomolecules and cellular structures. This loss of function can lead to physiological decline and neurodegeneration, such as in diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and, potentially, Huntington’s disease. The following experiments use the model ...


Molecular Switch Model For Cardiomyocyte Proliferation, Satwat Hashmi, H R. Ahmad Jan 2019

Molecular Switch Model For Cardiomyocyte Proliferation, Satwat Hashmi, H R. Ahmad

Department of Biological & Biomedical Sciences

This review deals with the human adult cardiomyocyte proliferation as a potential source for heart repair after injury. The mechanism to regain the proliferative capacity of adult cardiomyocytes is a challenge. However, recent studies are promising in showing that the 'locked' cell cycle of adult cardiomyocytes could be released through modulation of cell cycle checkpoints. In support of this are the signaling pathways of Notch, Hippo, Wnt, Akt and Jak/Stat that facilitate or inhibit the transition at cell cycle checkpoints. Cyclins and cyclin dependant kinases (CDKs) facilitate this transition which in turn is regulated by inhibitory action of pocket ...


Cardiac Effects Of Huntington’S Disease In Murine Models, Daniel David Child Jan 2019

Cardiac Effects Of Huntington’S Disease In Murine Models, Daniel David Child

Publicly Accessible Penn Dissertations

Huntington’s disease (HD) is an autosomal dominant disease known for its severe neurologic phenotype. However, the causative protein in HD, mutant huntingtin (mHTT), is widely expressed across tissue types and may have profound consequences for peripheral organs. Notably, heart disease is the second leading cause of death in HD patients, but if or how cardiac mHTT expression causes pathology is still unknown. Here, I characterize the cardiac phenotype in two HD mouse models and show that dysregulated mTORC1 activity is a key underlying mechanism. I show that normal heart growth is limited in HD mouse models, with mass regulated ...


Unraveling The Beta Cell Translatome: Elucidation Of An Erk/Hnrnpk/Jund Axis, Austin Lewis Good Jan 2019

Unraveling The Beta Cell Translatome: Elucidation Of An Erk/Hnrnpk/Jund Axis, Austin Lewis Good

Publicly Accessible Penn Dissertations

In type 2 diabetes, oxidative stress contributes to the dysfunction and loss of pancreatic β cells. A highly conserved feature of the cellular response to stress is the regulation of mRNA translation, however, the mechanisms underlying this process in β cells are not fully understood. Here we use TRAP-seq to examine changes in the ribosome occupancy of mRNAs during conditions associated with β cell dysfunction, leading us to identify a cohort of translationally regulated genes with 3’UTR enrichment of a cytosine-rich motif. Of particular interest was the gene encoding JUND, a transcription factor with anti-oxidant functions in other cell ...


Vitamin D Receptor Gene Polymorphisms Knowledge And Breast Cancer In Texas, Ejike Roland Egwuekwe Jan 2019

Vitamin D Receptor Gene Polymorphisms Knowledge And Breast Cancer In Texas, Ejike Roland Egwuekwe

Walden Dissertations and Doctoral Studies

Breast cancer is a world health problem and is a leading cause of cancer-related death among women in the United States. However, breast cancer risks were reported to be reduced through exposure to Vitamin D through its Receptors identified as the p53 target gene. The purpose of this study was to assess the associations between VDR gene polymorphisms knowledge/awareness and decisions to reduce breast cancer risks and likelihood of mammogram screening among women in Texas. Data from survey were used. Roy adaptation model was the theoretical framework that guided this quasi- experimental, quantitative research. The dependent variables were decisions ...


Identification Of Deubiquitinating Enzymes That Control The Cell Cycle In Saccharomyces Cerevisiae, Claudine E. Mapa Nov 2018

Identification Of Deubiquitinating Enzymes That Control The Cell Cycle In Saccharomyces Cerevisiae, Claudine E. Mapa

GSBS Dissertations and Theses

A large fraction of the proteome displays cell cycle-dependent expression, which is important for cells to accurately grow and divide. Cyclical protein expression requires protein degradation via the ubiquitin proteasome system (UPS), and several ubiquitin ligases (E3) have established roles in this regulation. Less is understood about the roles of deubiquitinating enzymes (DUB), which antagonize E3 activity. A few DUBs have been shown to interact with and deubiquitinate cell cycle-regulatory E3s and their protein substrates, suggesting DUBs play key roles in cell cycle control. However, in vitro studies and characterization of individual DUB deletion strains in yeast suggest that these ...


Stochastic Difference Model For Evolutional Dynamics Of Large Antigen Repertoires In African Trypanosomes, Fan Yu Oct 2018

Stochastic Difference Model For Evolutional Dynamics Of Large Antigen Repertoires In African Trypanosomes, Fan Yu

Annual Symposium on Biomathematics and Ecology: Education and Research

No abstract provided.


Ire1b Degrades Rnas Encoding Proteins That Interfere With The Induction Of Autophagy By Er Stress In Arabidopsis Thaliana, Yan Bao, Yunting Pu, Xiang Yu, Brian D. Gregory, Renu Srivastava, Stephen H. Howell, Diane C. Bassham Aug 2018

Ire1b Degrades Rnas Encoding Proteins That Interfere With The Induction Of Autophagy By Er Stress In Arabidopsis Thaliana, Yan Bao, Yunting Pu, Xiang Yu, Brian D. Gregory, Renu Srivastava, Stephen H. Howell, Diane C. Bassham

Genetics, Development and Cell Biology Publications

Macroautophagy/autophagy is a conserved process in eukaryotes that contributes to cell survival in response to stress. Previously, we found that endoplasmic reticulum (ER) stress induces autophagy in plants via a pathway dependent upon AT5G24360/IRE1B (INOSITOL REQUIRING 1–1), an ER membrane-anchored factor involved in the splicing of AT1G42990/BZIP60 (basic leucine zipper protein 60) mRNA. IRE1B is a dual protein kinase and ribonuclease, and here we determined the involvement of the protein kinase catalytic domain, nucleotide binding and RNase domains of IRE1B in activating autophagy. We found that the nucleotide binding and RNase activity of IRE1B, but not ...


The Role Of Mesenchymal Stromal Cells And Classical Dendritic Cells In The Maintenance And Regulation Of The Bone Marrow Niche, Jingzhu Zhang Aug 2018

The Role Of Mesenchymal Stromal Cells And Classical Dendritic Cells In The Maintenance And Regulation Of The Bone Marrow Niche, Jingzhu Zhang

Arts & Sciences Electronic Theses and Dissertations

The bone marrow niche is an important microenvironment for the regulation of normal and malignant hematopoiesis. The first discovered niche component is mesenchymal stromal cells, which are the major source for the production and secretion of multiple niche factors. Mesenchymal stromal cells are heterogeneous and various transgenes have been used to target non-identical but overlapping subpopulations. To further characterize the heterogeneity of mesenchymal stromal cells, we tested the targeting specificity of three tissue-specific Cre-recombinase transgenes. We show that in addition to osteoblasts, Ocn-Cre targets a majority of Cxcl12-abundant reticular (CAR) cells and arteriolar pericytes. Surprisingly, Dmp1-Cre also targets a subset ...


Understanding Nuage-Mitochondrial Coupling In Drosophila Pirna Biogenesis, Tianfang Ge Aug 2018

Understanding Nuage-Mitochondrial Coupling In Drosophila Pirna Biogenesis, Tianfang Ge

GSBS Dissertations and Theses

In the Drosophilaovary, PIWI-interacting RNAs (piRNAs) suppress transposon expression, ensuring female fertility. PIWI proteins Aub and Ago3, loaded with ping-pong piRNAs and reside in perinuclear nuage granules, engage in reciprocal transposon transcript cleavage termed the ping-pong cycle. The other PIWI protein Piwi, loaded with phased piRNAs and resides in the nucleus, silences transposon transcriptionally. Ping-pong piRNAs are made through the ping-pong amplification loop by Aub and Ago3, whereas phased piRNAs are made through consecutive endonucleolytic cleavages that spread in 5′-to-3′ direction, presumably by Zucchini (Zuc), an endonuclease resides on the surface of mitochondria. The ping-pong and phasing biogenesis ...


Effects Of The Everyday Toxin, Titanium Dioxide, On Drosophila Melanogaster Nervous System Development, Maria Carparelli, Kelsey Ripley, Stefanie Otto-Hitt Jul 2018

Effects Of The Everyday Toxin, Titanium Dioxide, On Drosophila Melanogaster Nervous System Development, Maria Carparelli, Kelsey Ripley, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

Titanium dioxide (TiO2) is a widely used compound found in everything from food packaging to sunscreens. When ingested, TiO2 is readily transported across membranes and efficiently stored within cells. Previous studies showed that exposure to TiO2 results in underdeveloped nervous systems. For our study, we attempted to answer the following question: Will exposing Drosophila melanogaster larvae to TiO2 affect expression of the Neur gene and development of the central nervous system? The Neur gene is crucial during the celldetermination stage of development as its encoded protein helps specify neuroblast development and aids in nervous system and sensory organ development. It ...


Effects Of Salt Concentration On Tetrahymena Thermophila Growth And Crp1 Gene Expression, Katie Mchugh, Rachel Tremaine, Anna Sapone, Stefanie Otto-Hitt Jul 2018

Effects Of Salt Concentration On Tetrahymena Thermophila Growth And Crp1 Gene Expression, Katie Mchugh, Rachel Tremaine, Anna Sapone, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The use of salt to remove ice from roads has resulted in increased salinity in many freshwater lakes in North America. For this project, we wanted to explore how environmental salt concentrations influence the rate of growth of Tetrahymena thermophila and expression of the CRP1 gene. We hypothesized that if the sodium concentration is increased in the media, Tetrahymena growth would decrease and expression of the CRP1 gene would increase. The CRP1 gene encodes a protein that helps regulate calcium concentrations within a cell based on the concentration of sodium ions. The media of the experimental group was treated with ...


Uv Light Exposure And Tetrahymena Thermophilia: Expression Of The Phr2 Gene, Molly Moloney, Tanner Gustavsen, Stefanie Otto-Hitt Jul 2018

Uv Light Exposure And Tetrahymena Thermophilia: Expression Of The Phr2 Gene, Molly Moloney, Tanner Gustavsen, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The PHR2 (photolyase repair) gene functions in the repair of DNA in Tetrahymena thermophila. It was hypothesized that when Tetrahymena cultures are exposed to UV light, increased expression of PHR2 would occur in response to the damage incurred. UV light has been known to cause the formation of pyrimidine dimers and other disruptions in DNA. Through two rounds of experimentation, the acute impact of UV light exposure was monitored. The cultures underwent RNA extraction, reverse transcription, and semi-quantitative PCR analysis. We predicted that UV exposure would result in the increased expression of PHR2.


Effect Of Potassium Bromate On Oxr1 Gene Expression And Cell Growth In Tetrahymena Thermophila, Scott Khale, Brendan Mcmahon, Stefanie Otto-Hitt Jul 2018

Effect Of Potassium Bromate On Oxr1 Gene Expression And Cell Growth In Tetrahymena Thermophila, Scott Khale, Brendan Mcmahon, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The purpose of our experiment was to answer the question: Does oxidative damage, induced by potassium bromate (KBrO3), affect growth rate and expression of the OXR1 gene in Tetrahymena thermophila? Potassium Bromate was chosen as the environmental stressor because it is a strong oxidizing agent that is used as a leavening agent in baked goods. It was hypothesized that expression of OXR1 would increase in Tetrahymena cells that were exposed to potassium bromate and that their growth rate would decrease. The exact mechanism and function of the OXR1 gene is still unknown; however, the literature suggests that it is required ...


Analysis Of Fbp1 Gene Expression In Uv Light-Exposed Tetrahymena Thermophila Cultures, Colby Henry, Stefanie Otto-Hitt Jul 2018

Analysis Of Fbp1 Gene Expression In Uv Light-Exposed Tetrahymena Thermophila Cultures, Colby Henry, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

In this study, Tetrahymena thermophila cultures were exposed to an acute treatment of UV light and expression of the FBP1 gene was examined. The FBP1 gene encodes Fructose- 1,6-biphosphatase which plays an important role in the process of gluconeogenesis. It was hypothesized that exposure of Tetrahymena to UV light would cause an upregulation in DNA repair enzymes, thereby causing these cells to require higher levels of glucose. The higher demand for glucose would then result in the upregulation of gluconeogenesis and FBP1. Following acute exposure of Tetrahymena cultures to UV light, the expression of FBP1 was analyzed using reverse ...


Effect Of Egta On Sit1 Scramblase Gene Expression And Cell Growth In Tetrahymena Thermophila, Emma Esposito, Theda Knauth, Amelia Ohnstad, Stefanie Otto-Hitt Jul 2018

Effect Of Egta On Sit1 Scramblase Gene Expression And Cell Growth In Tetrahymena Thermophila, Emma Esposito, Theda Knauth, Amelia Ohnstad, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

Scramblase is an enzyme that facilitates the movement of newly synthesized phospholipids from the cytosolic side to the extracellular side of the lipid bilayer. This process is vital for cell membrane repair and growth. In Tetrahymena thermophila, the gene SIT1 encodes for the Scramblase protein, whose functionality is Ca2+-dependent. In this experiment, the concentration of accessible Ca2+ ions was decreased in order to observe whether the change had an aect on the expression of SIT1 and cell growth. It was hypothesized that expression of the SIT1 gene would increase, while cell growth would decrease. To carry out the experiment ...


Effect Of Alcohol On Pfk1 Gene Expression And Feeding Activity In Tetrahymena Thermophila, Evelyn Sowers, Alex Skoulis, Stefanie Otto-Hitt Jul 2018

Effect Of Alcohol On Pfk1 Gene Expression And Feeding Activity In Tetrahymena Thermophila, Evelyn Sowers, Alex Skoulis, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The purpose of this experiment was to test whether alcohol induces starvation conditions in Tetrahymena thermophila. For this experiment, it was hypothesized that alcohol induces starvation due to a lack of energy available in the organism for feeding. This hypothesis was tested by monitoring food vacuole formation and expression of the PFK1 gene in Tetrahymena that were exposed to alcohol. The PFK-1 gene was chosen because its encoded protein plays an essential role in cellular metabolism. For the experiment, control and experimental cultures of Tetrahymena themophila were maintained in a nutrient rich media, with the media of the experimental group ...


The Effect Of Metal Pollutants On Cell Growth And Ttherm0058490expressionint Etrahymenathermophila, Inderbir Bains, William French, Stefanie Otto-Hitt Jul 2018

The Effect Of Metal Pollutants On Cell Growth And Ttherm0058490expressionint Etrahymenathermophila, Inderbir Bains, William French, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

In this experiment, we sought to answer the following question: Do higher levels of metal exposure affect expression of the TTHERM 00584900 gene in Tetrahymena thermophila? The TTHERM 00584900 gene produces an ATP-binding cassette protein that is responsible for removing metals and metalloids from Tetrahymena thermophila. It is hypothesized that the expression of TTHERM 00584900 will increase with exposure to metals. In order to test this hypothesis, Tetrahymena thermophila were randomly assigned to either a control group, which was cultured in a normal growth medium, or a treatment group, whichin addition to the medium was exposed to water contaminated with ...


Impacts Of Environmental Motion On Cdc2 Expression And Cell Growth In Tetrahymena Thermophila, Jack Rees, Keaton Brady, Stefanie Otto-Hitt Jul 2018

Impacts Of Environmental Motion On Cdc2 Expression And Cell Growth In Tetrahymena Thermophila, Jack Rees, Keaton Brady, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

In this experiment, the eect of constant motion on expression of the cell cycle control gene CDC2 was examined in the organism Tetrahymena thermophila. We hypothesized that CDC2 expression would decrease with a corresponding increase in movement within the Tetrahymena culture environment. The CDC2 protein is primarily used in the replication of eukaryotic DNA because of its connection to HHO1, an important factor in transcriptional regulation. If the cells were subject to vegetative growth in which there are no stressors put on the cell, then HHO1 becomes phosphorylated to prevent it from interacting with chromatin. Because CDC2 is important in ...


Effects Of Fatty Acids From Avocados On Expression Of The Hmg1 Gene In Tetrahymena Thermophila, Jasmine Phan, Connor Kane, Stefanie Otto-Hitt Jul 2018

Effects Of Fatty Acids From Avocados On Expression Of The Hmg1 Gene In Tetrahymena Thermophila, Jasmine Phan, Connor Kane, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The enzyme 3-Hydroxy-3-Methyl-Glutaryl-CoA Reductase (HMG) HMG catalyzes the ratelimiting step in the biosynthesis of cholesterol and is a common drug target for cholesterol reduction. Previous studies in rats have shown that avocado oil reduces cholesterol levels. We were interested in determining whether the healthy fatty acids in avocado oil would have an effect on the expression of HMG. In this study, Tetrahymena thermophila were treated with avocado oil to determine the effect of fatty acids on expression of the HMG1 gene that encodes HMG. Expression of HMG1 was monitored using reverse transcription and semi-quantitative PCR. We predicted that expression of ...


Exploring The Effects Of Environmental Temperature On Cdc-7 Expression In T. Thermophila, John Brothers, Alexis Gerry, Lauren Scofield, Stefanie Otto-Hitt Jul 2018

Exploring The Effects Of Environmental Temperature On Cdc-7 Expression In T. Thermophila, John Brothers, Alexis Gerry, Lauren Scofield, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

Tetrahymena thermophila are unicellular eukaryotes with a remarkably large genome: over 27,000 genes. One important aspect of studying the Tetrahymena genome is identifying and understanding what factors influence gene expression. This study examined the effects of temperature on the expression of CDC-7, a cell division control gene. We subjected Tetrahymena cultures to increased temperature and hypothesized such treatments would induce cell division and increase CDC-7 expression. Gene expression levels were measured via semi-quantitative PCR analysis after an acute heat treatment.


The Effect Of Basic Conditions On Cdk1 Expression In Tetrahymena Thermophila, Kaitlin Stromberg, Jakob Plagenz, Stefanie Otto-Hitt Jul 2018

The Effect Of Basic Conditions On Cdk1 Expression In Tetrahymena Thermophila, Kaitlin Stromberg, Jakob Plagenz, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The goal of this project was to discover whether expression of the CDK1 gene in Tetrahymena thermophila, as well as their growth, are aected by changing the pH of the growth media. We hypothesized that expression of the CDK1 gene would decrease significantly when grown in basic media. The CDK1 gene encodes the protein Cyclin-Dependent Kinase, which functions to advance cells through various stages of the cell cycle. To test our hypothesis, Tetrahymena cultures were randomly assigned to either the control group, which was grown in media with a neutral pH, or a treatment group, which was grown in media ...


The Effect Of Exposure To Uv Light On Rad51 Expression In Tetrahymena Thermophila, Kendall Patch, Kerri Mcinnis, Stefanie Otto-Hitt Jul 2018

The Effect Of Exposure To Uv Light On Rad51 Expression In Tetrahymena Thermophila, Kendall Patch, Kerri Mcinnis, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

In this experiment, we attempted to answer the question: Does exposure to UV light affect expressionof the Rad51 gene in Tetrahymena thermophila? It was hypothesized that the expression of Rad51 would increase proportionately to the exposure of UV light. The Rad51 protein is fundamental in the repair of double-stranded DNA breaks in eukaryotes, specifically during the process of homologous recombination. In order to test this hypothesis, Tetrahymena were randomly assigned to either a control group, which was cultured under ideal conditions, or a treatment group, which was exposed to UV light. Previous experiments have suggested that T. thermophila prosper most ...


Analyzing Atp Synthetase F1 Expression In Heat-Exposed Tetrahymena Thermophila, Kevin Gardner, Ian Lorang, Stefanie Otto-Hitt Jul 2018

Analyzing Atp Synthetase F1 Expression In Heat-Exposed Tetrahymena Thermophila, Kevin Gardner, Ian Lorang, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

In this study, we investigated the effect of temperature on expression of the ATP1 gene in the organism Tetrahymena thermophila. We hypothesized that expression of the ATP1 gene would increase as we increased temperature. ATP1 is a highly conserved gene in Tetrahymena that codes for the alpha subunit of ATP Synthase, an enzyme that generates the energy molecule ATP. We decided to use heat treatment based on studies performed on ATP1 in Alaskan Cod showing increased expression in response to increased temperatures (Michael et al., 2015). We used reverse transcription and semi-quantitative PCR to analyze expression of ATP1 in our ...


Effect Of Nitrate Pollution On Tetrahymena Thermophila, Reegan Debruycker, Hannah Sylvester, Stefanie Otto-Hitt Jul 2018

Effect Of Nitrate Pollution On Tetrahymena Thermophila, Reegan Debruycker, Hannah Sylvester, Stefanie Otto-Hitt

Life and Environmental Sciences Course-based Research Projects

The question under investigation in this project was: Does an elevated concentration of nitrates have a negative eect on cell growth and expression of the CDC16 gene in the organism Tetrahymena thermophila? Exposure of Tetrahymena cultures to nitrates mimics the exposure of these organisms to nitrate pollution in the environment, which can occur due to septic tank leakage, nitrogen-rich fertilizer run o, and agricultural processes. The EPA standard for nitrate concentrations in drinking water is a maximum of 10ppm. In our experiment, Tetrahymena cultures were treated with media that contained 30ppm of nitrates for one week. The 30ppm nitrate concentration ...