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Honors Theses

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Articles 31 - 60 of 103

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Elucidating The Interplay Between Sodium Selenite On The Tick Amblyomma Maculatum Selenoprotein Gene Expression, Afnan M. Beauti May 2017

Elucidating The Interplay Between Sodium Selenite On The Tick Amblyomma Maculatum Selenoprotein Gene Expression, Afnan M. Beauti

Honors Theses

Selenium (Se) is an element recognized as an essential micronutrient in eukaryote organisms. Selenoproteins contain selenium as selenocysteine, the 21st amino acid. Selenium plays a role in cell growth and functioning. At low concentrations, it can induce growth and at high concentrations, it can cause a cell to stop growing and potentially have toxic effects on the cell and organism. When selenium levels are high, oxidative stress results by the production of reactive oxidative species. Selenoproteins, however, can aid the antioxidant response in the cell. Ticks are arthropods of interest, as they are one of few that contain many selenogenes ...


Generation Of Mutant Snap-23 To Arrest Mast Cell Degranulation At Trans-Snare Complex Formation, Suzette Wafford-Turner May 2017

Generation Of Mutant Snap-23 To Arrest Mast Cell Degranulation At Trans-Snare Complex Formation, Suzette Wafford-Turner

Honors Theses

Regulated secretion of pro-inflammatory molecules (e.g, histamines, proteases) from mast cells plays critical roles in immunity, allergic reactions, cardiovascular disease and cancer. These molecules are stored in secretory granules inside the cell and are rapidly released into the extracellular environment when mast cells are activated. It is known that mast cell degranulation depends upon membrane anchored SNAREs (soluble N-ethylmaleidimide-sensitive factor attachment protein receptors) and accessory proteins that form the trans-SNARE complex, a 4 helical bundle central to exocytic fusion. There are three SNARE proteins that contribute to the 4-helical bundle during exocytosis; Syntaxin and VAMP proteins each provide one ...


Molecular Mass Of L-Arginine-Based Potentially Carcinogenic Heterocyclic Amines, Irene S. Hwang Apr 2017

Molecular Mass Of L-Arginine-Based Potentially Carcinogenic Heterocyclic Amines, Irene S. Hwang

Honors Theses

Investigation of cancer-inducing molecules in cooked foods has led to the discovery of mutagenic heterocyclic amines (HCAs) in meat. The amino acids creatin(in)e and L-phenylalanine form the precursors for these HCAs. Recent research here at Andrews University have found similar mutagenic HCAs produced from plant-based proteins when L-arginine, rather than creatin(in)e, is substituted in high temperature reactions (simulated cooking) with L-phenylalanine. Our research focused on developing HPLC methods to isolate individual candidates that are then screened for mutagenicity via the Ames test. Mass spectroscopy methods were also developed to identfy the molecular structure of these lead ...


Fluorescence Analysis Of Pamam Dendrimers, Ansel Nam Apr 2017

Fluorescence Analysis Of Pamam Dendrimers, Ansel Nam

Honors Theses

Despite their lack of conjugation, polyamidoamine (PAMAM) dendrimers are intrinsically fluorescent, and this fluorescent is somewhat manipulable through altering the dendrimer's solvent. This level of fluorescence is dependent on a number of the dendrimer's qualities: the pH of the solvent, the mass percent concentration of the dendrimer in solvent, and the generation size of the dendrimer. In past studies, fluorescent intensity increased as generation size increased and pH decreased. Thus, this research project will attempt to replicate those results as well as further analyze the efficiency of fluorescence of PAMAM dendrimer in comparison to fluorescein.


Vesicle-Membrane Tethering: The Interaction Of Sec1 And Exocyst Subunit Sec6, Lauren O'Connor Jan 2017

Vesicle-Membrane Tethering: The Interaction Of Sec1 And Exocyst Subunit Sec6, Lauren O'Connor

Honors Theses

The exocyst complex is a multi-subunit tethering complex that is used in the process of exocytosis. There are eight subunits in the complex, and these subunits interact with each other as well as proteins outside of the complex to facilitate membrane fusion. Prior research has shown that the exocyst subunit Sec6 and Sec1 from the Sec1/Munc18 (SM) family, a known regulator of membrane fusion, interact with each other. In this experiment, the goal was to crosslink the two proteins and in turn work to identify the interacting amino acid residues that are responsible for the proteins’ interaction. To do ...


Examining Interactions Between Opsins And Carotenoid Biosynthetic Proteins In Halophilic Archaea, Alexandru M. Plesa Jan 2017

Examining Interactions Between Opsins And Carotenoid Biosynthetic Proteins In Halophilic Archaea, Alexandru M. Plesa

Honors Theses

Organisms have evolved numerous specialized molecules for constantly responding to environmental changes. Examples of such molecules are the light-driven proton-pump rhodopsins, such as bacteriorhodopsin (BR) and cruxrhodopsin (cR), and the carotenoid pigments, such as retinal and bacterioruberin. In halophilic Archaea, retinal can covalently bind bacterioopsin (BO) and cruxopsin (CO) to form the corresponding protein complexes, and its biosynthesis is indirectly controlled by the activity of the lycopene elongase (Lye) enzyme, which converts lycopene, a retinal precursor, to a form of bacterioruberin. Interestingly, opsins were shown to inhibit the activity of Lye, thereby promoting retinal biosynthesis and indirectly regulating the apoprotein-cofactor ...


Investigating Medicinally Important Portein-Protein And Protein-Ligand Interactions : A Computational Approach, Cooper Ashley Taylor Jan 2017

Investigating Medicinally Important Portein-Protein And Protein-Ligand Interactions : A Computational Approach, Cooper Ashley Taylor

Honors Theses

Molecular dynamics (MD) simulations and computational chemistry allow for an atomistic understanding of protein-protein and protein-ligand binding motifs. Through the use of MD, medicinally relevant complexes can be examined in detail unattainable by experimental methods. Within this work, systems pertinent to both Alzheimer’s Disease and HIV-1 are probed and thoroughly sampled to help elucidate potential therapeutic pathways. We used molecular dynamics and free energy estimations to gauge the affinity for the binary and ternary complexes of KLC1, APP and JIP1, three proteins all believed to be involved in the propagation of Alzheimer’s Disease. Two areas of thought exist ...


Expression Of Glycine-Rich Proteins Found In Salivary Glands Of The Lone Star Tick (Amblyomma Americanum) Using A Mammalian Cell Line, Annabelle Clark Dec 2016

Expression Of Glycine-Rich Proteins Found In Salivary Glands Of The Lone Star Tick (Amblyomma Americanum) Using A Mammalian Cell Line, Annabelle Clark

Honors Theses

Ticks play an important ecological role as well as a growing role in human health and veterinary care. Ticks are hosts to a plethora of microbial pathogens that can be transferred during feeding to cause tick-borne diseases in humans and many animals. Ticks may in large part owe the success of the transfer of these pathogens between hosts to their complex saliva. The saliva secreted upon a tick’s attachment to a host serves the following, among other, functions: anti-hemostasis of the blood pool, preventing an inflammatory response at the bite site, and serving as a natural anti-microbial substance. An ...


Functional Significance Of Branch Points In Mirtrons, Britton A. Strickland Dec 2016

Functional Significance Of Branch Points In Mirtrons, Britton A. Strickland

Honors Theses

MicroRNAs are a heterogeneous group of small regulatory RNAs generated by many pathways. Mirtrons (miR) are a class of microRNAs produced by splicing, and some mirtrons contain a 3’ tail located downstream from the self-complementary hairpin. During RNA splicing, a loop-like “lariat” intermediate structure is created when the 5’ end of the RNA is attached to an adenine called the branch point. The goal of this project is to uncover the contribution of branch point location to the processing of tailed mirtrons into functional gene regulators. This project approaches this issue from two directions. First, branch points were identified by ...


Investigation Of The Caveolin-Human Follicle Stimulating Hormone Receptor Interaction Through Peptide Treatments, Timothy James Hagan Jun 2016

Investigation Of The Caveolin-Human Follicle Stimulating Hormone Receptor Interaction Through Peptide Treatments, Timothy James Hagan

Honors Theses

The purpose of my research project has been to determine the nature of the binding relationship between caveolin and hFSHR inside sex cells. FSH plays a role in the maturation of these cells, and interfering with the receptor’s interaction with caveolin would prevent cell maturation (down regulating fertility). It is believed the interaction occurs through transmembrane domain IV of the receptor due to its aromatic nature. The treatment of sex cells with synthetic peptides that mimic the hFSHR-caveolin binding sequence should prevent the interaction, shutting down the signaling cascade from hFSHR. This can be tested for by the monitoring ...


Tubulin Post-Translational Modifications Are Altered By Changes In Actin-Myosin Contractility In Non-Muscle Cells, James Mcgee Jun 2016

Tubulin Post-Translational Modifications Are Altered By Changes In Actin-Myosin Contractility In Non-Muscle Cells, James Mcgee

Honors Theses

All cells regulate their contractility by the interaction of actin and myosin. In non-muscle cells, however, this interaction is regulated by rho, a GTPase, which is the upstream effector for multiple pathways. Cell contractility increases when rho is active, and prior research has shown the depolymerization of microtubules (MTs) activates rho. Microtubules play a role in many cellular functions and it is thought that either associations with MT-associated proteins and/or post-translational modifications to tubulin regulate MT functions. Numerous post-translational modifications (PTMs) to microtubules have been identified, but their roles are not well understood. A possible relationship between changes in ...


Development Of A [2]-Catenane Synthetic Method And A Student Beliefs Survey For A Hybrid Organometallics Course, Jourdan E. Lakes May 2016

Development Of A [2]-Catenane Synthetic Method And A Student Beliefs Survey For A Hybrid Organometallics Course, Jourdan E. Lakes

Honors Theses

In this study, the value-added of a new upper-level chemistry course, CHEM 372-Organometallics, was defined using a Student Beliefs Survey that was answered by students in CHEM 372 and by faculty who teach upper-level chemistry courses at Coastal Carolina University. The laboratory portion of CHEM 372 focused on the synthesis of a [2]-catenane, which is a molecularly interlocked molecule (MIM). The intention was to give the CHEM 372 students a goal to work toward over the course of the semester instead of different, unrelated, experiments each week. Using this style of lab teaching, the students were introduced to graduate ...


Painless Hematophagy: The Functional Role Of Novel Tick Metalloproteases In Pain Suppression, Joseph W. Jelinski May 2016

Painless Hematophagy: The Functional Role Of Novel Tick Metalloproteases In Pain Suppression, Joseph W. Jelinski

Honors Theses

Ticks secrete a plethora of pharmacologically active molecules in their saliva while feeding. These allow the tick to feed upon a host over prolonged periods of time in an itch free and painless attachment. The exact mechanism of pain suppression by the tick has barely been investigated. In this study, two angiotensin converting enzymes (ACEs), members of the metalloprotease family, are identified as potentially responsible for the degradation of pro-inflammatory peptides. It is hypothesized that these tick ACEs block the function of bradykinin through degradation of the peptide, contributing to the tick’s ability to conduct a pain and itch-free ...


Using Intrinsic Properties Of Polyaniline To Sense Expression Of The Microrna Let-7, Jared N. Gloria May 2016

Using Intrinsic Properties Of Polyaniline To Sense Expression Of The Microrna Let-7, Jared N. Gloria

Honors Theses

MicroRNAs are approximately 22-nucleotide long RNA molecules that function through decay and translational repression of messenger RNA. The microRNA let-7 is found to play a role in maintaining the fate of differentiated cells in humans. Thus, expression level of this microRNA is a reliable biomarker of tumor cell phenotype. However, there are significant limitations in the current profiling techniques of microRNA. The current methods like northern blotting, microarrays, RT-PCR, or using locked nucleic acid (LNA) for in-situ hybridization are either laborious, semi-quantitative, or expensive. In this research we try to address this issue by developing a fast, specific, and inexpensive ...


Developing A Deletion Construct Of The Halothiobacillus Neapolitanus Csos1c Gene, Ellyn K. Dunbar May 2016

Developing A Deletion Construct Of The Halothiobacillus Neapolitanus Csos1c Gene, Ellyn K. Dunbar

Honors Theses

The purpose of this research was to develop a deletion construct for the chemoautotrophic bacterium Halothiobacillus neapolitanus, which will be used to generate a mutant lacking a carboxysome shell protein gene. The carboxysome is the location of carbon dioxide fixation. The operon that encodes the carboxysome contains three genes for CsoS1 proteins, the major components of the carboxysome shell. The small CsoS1 proteins self-assemble into hexamers with small central pores. The hexamers arrange into the facets of the icosahedral carboxysome shell. The pores are believed to be involved in selective diffusion of materials necessary for carbon dioxide fixation across the ...


Effects Of Creatine, Leucine, And Ethanol On Glial Cell Line-Derived Neurotrophic Factor In C2c12 Skeletal Muscle Cells, Daniel Waters Apr 2016

Effects Of Creatine, Leucine, And Ethanol On Glial Cell Line-Derived Neurotrophic Factor In C2c12 Skeletal Muscle Cells, Daniel Waters

Honors Theses

Glial Cell Line-Derived Neurotrophic Factor (GDNF) is a protein expressed throughout the mammalian nervous system that serves several protective functions. Originally discovered in the brain, where it supports the survival of dopaminergic neurons, GDNF has since been shown to be expressed in skeletal muscle and is vital for the maintenance of motor neurons. The relationship between muscle hypertrophy, and GDNF is not well understood. The purpose of this study was to examine the relationship between hypertrophy and GDNF by inducing hypertrophy in C2C12 skeletal muscle cells using leucine and creatine, inhibiting hypertrophy using ethanol and measuring GDNF content. As measured ...


Determining The Concentration Of Nitric Oxide In Solvent, Emily-Jean E. Bankes Mar 2016

Determining The Concentration Of Nitric Oxide In Solvent, Emily-Jean E. Bankes

Honors Theses

The body creates nitric oxide (NO) for signaling. One way that researchers study NO signaling is through the use of model complexes, or molecules that have the same metal ion and a similar molecular shape as biological molecules and are easy to synthesize. Currently researchers have limited ways to deliver precisely-known small quantities of NO to an experiment where they are trying to investigate NO-binding to a transition metal complex. A peak shift in the absorption spectrum at ~530 nm makes it possible for UV-Vis spectroscopy to observe NO binding to cobalt tetraphenylporphyrin (CoTPP). This makes it possible to measure ...


Discovering New Antibiotics: Bacterial Extracts Separated By Thin-Layer Chromatography Inhibit The Growth Of Staphylococcus, Heidi Hughes Jan 2016

Discovering New Antibiotics: Bacterial Extracts Separated By Thin-Layer Chromatography Inhibit The Growth Of Staphylococcus, Heidi Hughes

Honors Theses

Many bacteria have become resistant to commonly used antibiotics because of antibiotic use in people and animals. Therefore, new antibiotics are needed that will inhibit these resistant bacteria. Bacteria found in soil are a likely source for new antibiotics because of the limited available nutrients found in the soil. We isolated soil bacteria and screened them for antibiotic production against Staphylococcus epidermidids. Methanol extracts were made from entire agar plates of the soil bacteria that inhibited S. epidermidis. These extracts were spotted on a lawn of Staphylococcus aureus; growth inhibition was measured to comfirm that the extracts contained the antimicrobial ...


Computational Studies Of Paradifluorobenzene Cations And Hydrogen Cyanide Molecule, John C. Rowe Iv Jan 2016

Computational Studies Of Paradifluorobenzene Cations And Hydrogen Cyanide Molecule, John C. Rowe Iv

Honors Theses

Polycyclic aromatic hydrocarbons (PAHs) are hypothesized to comprise a significant portion of interstellar carbon identified from the Diffuse Interstellar Bands (DIBs). Gas phase cation-molecule reactions between compounds that previously existed as weakly bound species provides the best explanation for the emergence of PAHs in the interstellar medium (ISM). In this work, we use computational methods to characterize one of these weakly-bound systems. We discovered that intramolecular charge distribution (calculated using the Natural Population Analysis) dictate the complexes formed between a paradifluorobenzene cation and either one or two HCN molecules. Additionally, the relative stabilities of the complexes and the binding energies ...


Characterization Of Catecholamine Receptors And Transporters In Murine Macrophages, Elizabeth Gonye Jan 2016

Characterization Of Catecholamine Receptors And Transporters In Murine Macrophages, Elizabeth Gonye

Honors Theses

Macrophages are a critical part of the immune response. When circulating monocytes move into tissues they differentiate int macrophages to mount the first line of defense against pathogens.


Development Of A Chemical Genetic Screen To Determine Synergistic Compounds With Laromustine In Treating Glioblastoma Multiforme Cultured Cells, Ryan Weeks Jan 2016

Development Of A Chemical Genetic Screen To Determine Synergistic Compounds With Laromustine In Treating Glioblastoma Multiforme Cultured Cells, Ryan Weeks

Honors Theses

Laromustine is a chemotherapeutic sulfonylhydrazine prodrug used in clinical trials to treat acute myeloid leukemia (AML) and glioblastoma multiforme (GBM). While treatment of AML with laromustine has more demonstrative clinical success, there are enough promising data against GBM to pursue additional pre-clinical and clinical experiments. To determine the synergistic effects caused by treating cultured GBM cells with laromustine and a library of FDA-approved compounds, a chemical genetic screen was developed. To optimize the screen, optimal cultured GBM cell seed density, growth period and maximum well capacity were determined. The treatment period for a lethal dose of laromustine in cultured GBM ...


Impact Of Vector Range Expansion On Pathogen Transmission Dynamics Of Lyme Disease In Southwestern Virginia, Bishan Bhattarai Jan 2016

Impact Of Vector Range Expansion On Pathogen Transmission Dynamics Of Lyme Disease In Southwestern Virginia, Bishan Bhattarai

Honors Theses

Blacklegged ticks, Ixodes scapularis is the primary vector of Lyme disease in eastern United States. Borrelia Burgdorfei, the etiological agent of Lyme disease is transferred by ticks of Ixodes species. In recent years, its congener, Ixodes affinis has been expanding its range northwards from its southern population. We were interested in studying how the introduction of this new vector affected the interaction between the pathogen genotype and the host. We hypothesized that differential host use by I. affinis and I. scapularis would partly explain observed differences in B. burgdorferi infection prevalence and genotypic structure in southeastern Virginia. The result from ...


Calcium (Ii) Catalyzed Nitrone Additions, Elizabeth A. Congdon Jan 2016

Calcium (Ii) Catalyzed Nitrone Additions, Elizabeth A. Congdon

Honors Theses

Calcium(II) complexes have be shown to be successful catalysts for nitrone reactions. The addition of n-­‐methyl and n-­‐phenyl nitrones to donor-­‐acceptor cyclopropanes was achieved with calcium triflate (Ca(OTf). Differentially substituted tetrahydro-­‐1,2-­‐oxazines were synthesized in good to excellent yields. Calcium triflate was also found to catalyze the addition of silyl enol ethers to n-­‐phenyl nitrones along the Mukaiyama-­‐Mannich addition pathway. B-­‐amino carbonyls were synthesized from a variety of substituted nitrones. Bulky and cyclic silyl enol ethers were also found to be reactive, the products of which were isolated in good ...


Granulins As Inflammatory Mediators In Alzheimer Disease, Randi G. Reed May 2015

Granulins As Inflammatory Mediators In Alzheimer Disease, Randi G. Reed

Honors Theses

Alzheimer disease (AD) is a neurodegenerative disorder characterized by severe memory deficit and cognitive decline among the elderly. This degeneration is caused by the aggregation and deposition of a protein called amyloid-β (Aβ) in the brain. Aggregation of Aβ causes neuroinflammation in addition to other toxic events. However, it is unclear whether inflammation from an external source, such as from a traumatic brain injury (TBI), could trigger Aβ aggregation. In this context, several pro-inflammatory mediators such as cytokines and chemokines have been suspects. It is now hypothesized that a group of proteins called granulins (Grns) are unique inflammatory mediators that ...


Using Golden Gate Assembly To Rapidly Create Genetic Constructs For Transgenic Plants And Promoter Analysis, Jonathan Smith Apr 2015

Using Golden Gate Assembly To Rapidly Create Genetic Constructs For Transgenic Plants And Promoter Analysis, Jonathan Smith

Honors Theses

Golden Gate Assembly, a genetic assembly method in which the ligation and digestion of oligonucleotides occurs in a single step, can be used for an array of purposes. In order to test and take advantage of this assembly method our lab selected three experiments in which it could be used. These included an oxidative stress sensitive green florescent protein, a plant specific form of Golden Gate Assembly coined "Green Gate Assembly," and a viral promoter verification system called "pClone Red."

Arabidopsis (Arabidopsis thaliana) plants were analyzed for the presence of a redox-sensing green fluorescent protein (reduction-oxidation-sensitive green fluorescent protein [roGFP ...


Identification Of Bacterial Species Using Colony Pcr, Kaiti Walker Apr 2015

Identification Of Bacterial Species Using Colony Pcr, Kaiti Walker

Honors Theses

My research consisted of two separate parts, both involving the PCR methods. In part one I was trying to identify bacteria isolated from a chicken coop. In part two I was trying to identify bacteria isolated from the bluff. In both parts, I used a technique called colony PCR. This concept uses a single colony of bacteria to complete the PCR reaction. One pipet tip from a single colony is selected from the plate and mixed into a tube containing ultrapure water. A small amount of this bacterial mixture is then added into the PCR reaction. In the first part ...


Fatty Acid Content In Shallow And Mesophotic Colonies Of The Scleractinian Corals Montastraea Cavernosa And Agaricia Agaricites From The Cayman Islands, Jessica Ciesla Jan 2015

Fatty Acid Content In Shallow And Mesophotic Colonies Of The Scleractinian Corals Montastraea Cavernosa And Agaricia Agaricites From The Cayman Islands, Jessica Ciesla

Honors Theses

Two Scleractinian corals, Agaricia agaricites and Montastraea cavernosa, were collected from shallow (20 m depth) and mesophotic (60 m) ecosystems from the Cayman Islands. The fatty acid content of these species was analyzed by gas chromatography to determine variation in fatty acid profiles between the species and within species at different depths (shallow and mesophotic). Fatty acid composition is indicative of an organism’s diet and can be used to determine feeding habits of corals. The corals studied had different fatty acid content, suggesting different feeding mechanisms. Additionally, a difference in fatty acid content arose as a result of living ...


Biochemical Conversion Of Biomass Into Butanol Using Clostridium Acetobutylicum, Eric Stevens Jan 2015

Biochemical Conversion Of Biomass Into Butanol Using Clostridium Acetobutylicum, Eric Stevens

Honors Theses

Butanol is a potential transportation fuel which could eventually replace fossil- derived petroleum and transition the world towards a more sustainable future. This fuel can be produced by enzymatically hydrolyzing pretreated lignocellulosic biomass, and simultaneously fermenting the resultant sugars with specific bacterial strains. In this experiment switchgrass was chemically pretreated in a 75% (v/v) ethanol and 1% H2SO4 (1% v/v) solution and subjected to simultaneous saccharification and fermentation (SSF) with hydrolytic enzymes and Clostridium acetobutylicum. The pH control method for the SSF apparatus was varied between a 50 mM acetate buffer solution (HOAc), and the addition of calcium ...


An Nmr-Guided Approach To The Isolation Of Secondary Metabolites From Nrps And Pks Gene Clusters, Connor Craig Jan 2015

An Nmr-Guided Approach To The Isolation Of Secondary Metabolites From Nrps And Pks Gene Clusters, Connor Craig

Honors Theses

Secondary metabolites continue to offer a promising source of novel chemical entities with therapeutic bioactivities. Our research aims to isolate secondary metabolites produced by non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) constructs. A metagenomic library was created from the marine bacterium Pseudoalteromonas sp., which was isolated from the Chesapeake Bay sponge Halichondria bowerbanki. Using a functional screen for PPTase activity we were able to identify colonies that contained NRPS and PKS gene clusters. Cultures of this microbe have been produced and analyzed using a combination of chromatography and 1H, 13C, HMBC, HSQC, COSY, AND NOESY NMR spectra. We were ...


Expression Analysis Of A Glucose Oxidase Transgene In Tobacco, Jordan Burt Jan 2015

Expression Analysis Of A Glucose Oxidase Transgene In Tobacco, Jordan Burt

Honors Theses

Glucose oxidase (GOX) is an enzyme in plants that catalyzes the oxidation of glucose to hydrogen peroxide and Glucono delta-lactone[5]. We have expressed GOX under the control of an estrogen inducible system, XVE, to analyze the gene's expression under this system compared to a system using the 35s system. The 35s system contains a promoter that constitutively turns on the GOX gene in the Nicotiana tabacum plant which causes the gene to always be turned on. Leaf disc assays were performed with discs from 35s, XVE, and also wild type plants (not containing the GOX gene) in order ...