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Full-Text Articles in Molecular Biology

Investigating The Role Of Dnc-2 And Dli-1 On Ampa Receptor Mediated Behaviors In Caenorhabditis Elegans, Natalie Oberding Apr 2018

Investigating The Role Of Dnc-2 And Dli-1 On Ampa Receptor Mediated Behaviors In Caenorhabditis Elegans, Natalie Oberding

Life and Environmental Sciences Undergraduate Theses

Γ-amino-3-hydroxy-5methylisoxazole-4-proprionic acid (AMPA) receptors (AMPARs) are protein complexes involved in excitatory neurotransmission. AMPARs are tetrameric structures consisting of the pairings of GluA1, GluA2, GluA3, and GluA4 subunits. Dctn2 and Dync1li1 were found to interact with GluA2 in an immunoprecipitation screen that was performed on postnatal day 14 rat brains. DNC-2 and DLI-1 were identified as the C. elegans homologs of Dctn2 and Dync1li1, respectively, and are components of motor proteins that transport cargo throughout the cell. The goal of this project was to determine whether knocking down DNC-2 and DLI-1 by RNAi affects AMPA Receptor-mediated behaviors, including mechanosensation and chemosensation ...


Is Hif-1Α Required For V-Atpase-Dependent Regulation Of Glycolytic Enzymes?, Tyler Northrup Apr 2016

Is Hif-1Α Required For V-Atpase-Dependent Regulation Of Glycolytic Enzymes?, Tyler Northrup

Life and Environmental Sciences Undergraduate Theses

Vacuolar-ATPase (V-ATPase) is frequently up-regulated in tumors and tumor cell lines where it contributes to tumorigenic phenotypes, including the Warburg Effect (e.g. the phenomenon where glycolysis is preferred over oxidative phosphorylation (Vander Heiden et al. 2009)). The preferential use of glycolytic metabolism favored by tumor cells may be caused by at least two factors (Vander Heiden et al. 2009). First, the metabolic intermediates generated by the citric acid (TCA) cycle are used in a variety of anabolic pathways in the cell, and may supply a ready source of intermediate buildingblocks for biosynthetic processes. Second, hypoxia observed in some tumors ...


Investigating The Role Of Prp And Amyloid Beta Proteins In Ad And Other Protein-Misfolding Diseases, Steven Cornish Apr 2015

Investigating The Role Of Prp And Amyloid Beta Proteins In Ad And Other Protein-Misfolding Diseases, Steven Cornish

Life and Environmental Sciences Undergraduate Theses

Prion diseases are very rare, neurodegenerative diseases caused by misfolding of the Prion protein. The pathologies created by the misfolded protein are remarkably similar to those of Alzheimer’s disease. This paper provides a comprehensive study of the genomic and proteomic similarities between Prion proteins and several proteins implicated in Alzheimer’s disease (AD). While genomic similarity has been proven, studies are unclear as to whether any genomic mutations in PRNP affect the progression of AD. Studies have definitively proven that Prion protein interacts on a molecular level with several proteins implicated in AD. These interactions require further study to ...


Effect Of Membrane Cholesterol Levels And Allelic Variation On Prion Conversion, Ross Hartman Apr 2010

Effect Of Membrane Cholesterol Levels And Allelic Variation On Prion Conversion, Ross Hartman

Life and Environmental Sciences Undergraduate Theses

Chronic Wasting Disease (CWD) is caused by an accumulation of misfolded prion proteins (PrPsc) and subsequent plaque formation in the central nervous system. CWD is horizontally transferable; misfolded prions from one animal can enter another and cause normal prion proteins (PrPc) to misfold. This misfolding process is termed prion conversion. In natural deer populations an allelic variation in the prion gene is thought to confer resistance to CWD. Wild type mule deer are serine (S) homozygotes at codon 225. Mule deer that are Serine/ Phenyalanine (S/F) heterozygotes exhibit resistance to prion infection. In this study the F encoding allele ...


The Effects Of Glycosylation On Prion Protein Conversion, Christopher Casebeer Apr 2010

The Effects Of Glycosylation On Prion Protein Conversion, Christopher Casebeer

Life and Environmental Sciences Undergraduate Theses

Glycosylation of the prion protein may be experimentally altered through expression in different insect cell lines. The presence or absence of several terminal sugars may or may not have an effect on the conversion efficiency of the prion protein into its infectious form. The main focus of this research was to characterize the prion protein as it is produced in two varying insect cell lines that are able to glycosylate proteins to different extents. Glycosidase treatment demonstrated that the expressed prion protein was glycosylated. Lectin blotting was used to assess the presence or absence of terminal sugar moieties existing on ...


The Development Of A Quantitative Assay To Measure The Effects Of Protein Disulfide Isomerase On Prion Protein Misfolding, Lauren Vogl Apr 2010

The Development Of A Quantitative Assay To Measure The Effects Of Protein Disulfide Isomerase On Prion Protein Misfolding, Lauren Vogl

Life and Environmental Sciences Undergraduate Theses

The objective ofthis project was to develop a quantitative assay to measure the misfolding ofprion proteins that can be used to assess the effects of Protein Disulfide Isomerase (PDI) on the misfolding process ofthe Chronic Wasting Disease (CWD) prion protein. CWD is a spongiform encephalopathy native to animals in the family Cervidae, which includes whitetail deer, mule deer, elk and moose. CWD is caused by the misfolding ofprion proteins. Qualitative observations from previous studies indicate that disulfide bond rearrangement might be a rate limiting step in the misfolding process. There is a need to be able to quantitate the amount ...


The Effect Of Protein Disulfide Isomerase (Pdi) On In Vitro Conversion Of Mule Deer Prion Proteins, Alexander Nixon Apr 2009

The Effect Of Protein Disulfide Isomerase (Pdi) On In Vitro Conversion Of Mule Deer Prion Proteins, Alexander Nixon

Life and Environmental Sciences Undergraduate Theses

The objective of this project was to study Chronic Wasting Disease (CWD), a form of spongiform encephalopathy native to animals in the family Cervidae. The infectious agent in CWD is a misfolded form of the prion protein (PrPSc), which has undergone conversion from the normal, cellular prion protein (PrPc). The prion protein contains a disulfide bond, and the current project examined whether disulfide bond rearrangement is involved in the misfolding process. Using Protein Disulfide Isomerase (PDI), an enzyme that accelerates disulfide rearrangement, the efficiency of prion protein misfolding was monitored. Two different sources of normal prion protein were used: deer ...


Developing A Method For In Vitro Conversion Of Recombinant Mule Deer Prion Protein, Andrew Peterson Apr 2009

Developing A Method For In Vitro Conversion Of Recombinant Mule Deer Prion Protein, Andrew Peterson

Life and Environmental Sciences Undergraduate Theses

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) occurring in free-ranging and captive populations of deer and elk. CWD is thought to involve a misfolded protein called the prion protein. The misfolding SC mechanism remains unknown, but the conversion results from misfolded PrP contacting normal PrPc, and inducing a conformational change of PrPcto PrPsc. Previous studies have shown the normal cellular PrPc, isolated from mule deer brain, has the ability to misfold in vitro when incubated with misfolded PrPsc. Instead of using deer brain as a source of the normal cellular form of the prion protein, this project ...


The Effect Of Culture Medium Composition On The Localization Of Peroxisomal Membrane Proteins In Yeast, Maria Miller Apr 2009

The Effect Of Culture Medium Composition On The Localization Of Peroxisomal Membrane Proteins In Yeast, Maria Miller

Life and Environmental Sciences Undergraduate Theses

Formation and degradation of peroxisomes are not well understood, but their absence leads to serious consequences. In the study described in this thesis, 1 sought to (1) verify previous work indicating that switching from glucose-rich medium to lipid-rich medium induces peroxisome formation; (2) reveal that switching back from lipid-rich to glucose-rich medium induces peroxisome degradation; (3) test the hypothesis that Pex3p is a class II PMP that enters the peroxisome from the endoplasmic reticulum during de novo peroxisome formation; and (4) test the hypothesis that Pxalp is a class I PMP that enters the peroxisomal membrane from the cytosol. I ...


Cloning Of A Rare Chronic Wasting Disease Prion Allele From Mule Deer, Martha Bankhead Apr 2009

Cloning Of A Rare Chronic Wasting Disease Prion Allele From Mule Deer, Martha Bankhead

Life and Environmental Sciences Undergraduate Theses

The prion protein is thought to be the causative agent for Chronic Wasting Disease in mule deer (Odocoileus hemionus). Prion protein polymorphisms at amino acid 225 cause three different allele product combinations: homozygous 225 serine/serine, heterozygous 225 serine/phenylalanine, and homozygous 225 phenylalanine/phenylalanine. The most common combination of allele products in clinically progressed CWD in O. hemionus is serine/serine at 225. The phenylalanine substitution is underrepresented in diseased animals. The goal of this project was to clone the very rare phenylalanine encoding prion allele for future study. The polymerase chain reaction (PCR) was used to amplify the ...


The Use Of Sm-Fret Spectroscopy To Determine Whether Cooperative Binding Is Involved In The Chaperone Function Of Hiv-1 Nucleocapsid Protein, Elizabeth Swanson Apr 2008

The Use Of Sm-Fret Spectroscopy To Determine Whether Cooperative Binding Is Involved In The Chaperone Function Of Hiv-1 Nucleocapsid Protein, Elizabeth Swanson

Life and Environmental Sciences Undergraduate Theses

Human immunodeficiency virus nucleocapsid protein (HIV-1 NC) is known to have both structural and nucleic acid chaperone functions in the replication cycle of the retrovirus. As a nucleic acid chaperone, NC protein interacts with TAR RNA and TAR DNA structures during the minus-strand transfer step of reverse transcription. The aim of this study is to use single molecule florescence resonance energy transfer (SM-FRET) spectroscopy to study biotin-immobilized TAR DNA hairpins at various concentrations of NC protein. The resulting data will subsequently be used to determine whether cooperative binding occurs between the NC protein and the TAR DNA hairpins. The results ...


Neurological Effects Of Chlorpyrifos On Prenatally-Exposed Rats, Kyle Denning Apr 2008

Neurological Effects Of Chlorpyrifos On Prenatally-Exposed Rats, Kyle Denning

Life and Environmental Sciences Undergraduate Theses

Though extensively documented, autism is still poorly understood. Possible factors include prenatal or neonatal chemical exposure that causes declined synaptic transmission. One chemical, the pesticide known as chlorpyrifos, has been shown to cause anatomical differences in the areas of rats’ brains that are linked to behavioral symptoms closely related to autism. With the knowledge of the increased susceptibility ofmales to prenatal drug exposure, it was hypothesized that male rats exposed to chlorpyrifos before birth would exhibit such symptoms. Two tests, an elevated plus maze and a Morris water maze, were used to test anxiety and cognition, respectively. Anxiety tests yielded ...


Prion Infection Of The Nasal Cavity: Site For Agent Shedding?, Brian Moreaux Apr 2006

Prion Infection Of The Nasal Cavity: Site For Agent Shedding?, Brian Moreaux

Life and Environmental Sciences Undergraduate Theses

The mechanism of prion shedding in prion disease is currently unknown. The purpose of this study was to determine if the nasal cavity is a site of prion infection. Hamsters were intracerebrally inoculated with the hyper strain of transmissible mink encephalopathy agent. Once clinically ill, the hamsters were euthanized and their skulls were removed and examined for prion infection using immunohistochemical and dual immunofluorescence staining techniques. In infected hamsters, the infectious agent (PrP50) was shown to be present in the olfactory receptor neurons of the olfactory sensory epithelium (OSE) as well as in the nasal-associated lymphoid tissue and the vomeronasal ...


Rescuing A Molecular Motor In Dilute Mice, Sara Anderson Apr 2006

Rescuing A Molecular Motor In Dilute Mice, Sara Anderson

Life and Environmental Sciences Undergraduate Theses

Molecular motors are cellular enzymes that convert chemical energy into mechanical energy and are vital to cellular organization and function. The focus of the following research is Myosin Va, a molecular motor whose activity contributes to melanosome distribution in melanocytes. A mutation within myosin Va leads to the dilute coat-color phenotype in mice and some cases of Griscelli’s syndrome in humans. The main goal of this research was to create a chimeric myosin V, with portions of both myosin Va and Vb. This chimeric myosin was then transfected into dilute melanocytes to see if the normal distribution ofmelanosomes was ...


Perturbation Of Triple-Helix Structure In Mutant Collagens Examined By Subnanosecond Local Dynamics Of Tryptophan: An Application Of Time-Resolved Fluorescence Anisotropy, R. Dawn Paul Apr 2005

Perturbation Of Triple-Helix Structure In Mutant Collagens Examined By Subnanosecond Local Dynamics Of Tryptophan: An Application Of Time-Resolved Fluorescence Anisotropy, R. Dawn Paul

Life and Environmental Sciences Undergraduate Theses

Time-resolved fluorescence anisotropy can provide information about the local dynamic properties of a molecule at sub-nanometer resolution on the nano- and picosecond time scales. To help explain perturbations of the triple-helical structure of collagen induced by a mutation, and to facilitate an understanding of mutations associated with diseases like scurvy, an examination of synthetic collagen-like peptides was conducted. Results indicate that the extent of local motion increases near a mutation, demonstrating that the change in the amino acid interrupts the helical structure. Local motion, however, remains unchanged several amino acid residues away from the indole side chain of the tryptophan ...


Toward Solving The Structure Of Cholesterol Oxidase H447e/E361q Double Mutant, Larryn Krause Apr 2004

Toward Solving The Structure Of Cholesterol Oxidase H447e/E361q Double Mutant, Larryn Krause

Life and Environmental Sciences Undergraduate Theses

Cholesterol oxidase, a bifunctional flavoenzyme facilitates the oxidation and isomerization of cholesterol. Wild type cholesterol oxidase has been crystallized, and its structure has been resolved to 0.95 A. Wild type cholesterol oxidase turns over cholesterol with such extreme efficiency that it is very difficult to crystallize the substrate-enzyme complex. Cholesterol oxidase H447E/E361Q double mutant, which binds cholesterol, but is unable to completely turn it over, was expressed, purified, and crystallized to obtain a crystal structure. Once solved, the structure of the cholesterol oxidase double mutant could then be used as a background structure for the crystallization of the ...


Effects Of Estradiol And Tamoxifen On Leukocyte Distribution In Response To Restraint Stress In Mice (Mus Musculus), Julianna Duchesneau Apr 2004

Effects Of Estradiol And Tamoxifen On Leukocyte Distribution In Response To Restraint Stress In Mice (Mus Musculus), Julianna Duchesneau

Life and Environmental Sciences Undergraduate Theses

Selective estrogen receptor modulators (SERMs), drugs that either mimic or block the action of estrogen but do not have its side effects, have been under intense research since the early 20th century because of their distinct biological functions as antagonists and agonists in cells through their unique molecular conformations. Previous, unpublished research has found a correlation between a reduction in the leukocytic stress response in mice and injection with a natural form of estrogen, estradiol. The present study tested the effects of a SERM, tamoxifen, on the leukocyte distribution in mice after subjection to restraint stress. No difference was found ...


The Effect Of Pkc 5 Inhibitors On Pma-Induced Apoptosis In Xenopus Laevis Splenocytes, Annie Crater Apr 2003

The Effect Of Pkc 5 Inhibitors On Pma-Induced Apoptosis In Xenopus Laevis Splenocytes, Annie Crater

Life and Environmental Sciences Undergraduate Theses

Protein kinase C (PKC) controls apoptosis in amphibian systems such as that of the South African clawed frog, Xenopus laevis. The effects of cancer inducing agents like phorbol 12-myristate 13-acetate (PMA) when combined with Ca2+ independent PKC inhibitors on programmed cell death were studied in A laevis splenocytes. Regulation of PMA-activated cells was observed via Annexin V-FITC and propidium iodide assays, flow cytometry, and Western blot analysis. Results indicate the PKC inhibitors reduced both apoptosis and cell growth in PMA-stimulated X. laevis splenocytes compared to PMA alone. These results are valuable to future mammalian cancer research. By identifying the various ...


Analysis Of Candidate Proteins Interacting With Eyal, Ryan Schwanke Apr 2002

Analysis Of Candidate Proteins Interacting With Eyal, Ryan Schwanke

Life and Environmental Sciences Undergraduate Theses

The Eyal gene is important for mammalian organogenesis, and mutations in the human EYA1 results in Branchio-Oto-Renal (BOR) syndrome. This syndrome is autosomal dominant and causes combinations of branchial, otic, and renal anomalies. All Eya gene products contain an extremely conserved 271 amino acid C-terminus that is essential in protein-protein interaction. Recently, it was found that during mouse development, the expression of certain Six genes relied on the function of Eyal, and that Eyal interacts with relevant Six proteins through its C-terminal domain. These results provided insight into the molecular and developmental basis for organ defects occurring in BOR syndrome ...


Immunoprecipitation Of A Nadph Oxidase System Gtpase, Rac2, From The Human Neutrophil And The Gtp-Dependent Activity Of Rac2 In A Phosphorylation-Dependent Cell-Free System, Rachel Brennan Apr 2001

Immunoprecipitation Of A Nadph Oxidase System Gtpase, Rac2, From The Human Neutrophil And The Gtp-Dependent Activity Of Rac2 In A Phosphorylation-Dependent Cell-Free System, Rachel Brennan

Life and Environmental Sciences Undergraduate Theses

The neutrophil is an integral part of the body’s line of defense against infectious microbial agents. Its microbicidal activity is carried out by NADPH oxidase, an enzyme that uses electron transfer to create superoxide (O2"), which is then transformed into various toxins that are responsible for the demise of the foreign microorganism. The oxidase system is composed of membrane and cytosolic components. One of the cytosolic components is a low molecular-weight member of the Rho family of GTPases, Rac2. A genetic link has recently been established between Rac2 and Chronic Granulomatomous Disease (CGD), a disease in which the body ...


The Involvement Of C-Terminal Amino Acids In The Formation Of Prion Protein Dimers, Rebecca Phillip Apr 2001

The Involvement Of C-Terminal Amino Acids In The Formation Of Prion Protein Dimers, Rebecca Phillip

Life and Environmental Sciences Undergraduate Theses

Accumulations o f abnormal prion protein are thought to be the cause o f the transmissible spongiform encephalopathies, which are degenerative brain diseases. Throughout the disease process it is hypothesized that there is some sort of binding that occurs among the proteins. This research project involved the search for a strong, most likely covalent, linkage in a dimer of prion proteins. The focus was on the amino acid sequence o f the prion protein from 220 to 232. The lysine residue in this sequence was of particular concern due to its involvement in covalent linkages in other proteins (Priola et ...


Studies Of Mosaicism For Chromosomal Aneuploidy In Fetal Loss Using Fluorescence In Situ Hybridization On Several Tissue Types, Aaron Knox Apr 1999

Studies Of Mosaicism For Chromosomal Aneuploidy In Fetal Loss Using Fluorescence In Situ Hybridization On Several Tissue Types, Aaron Knox

Life and Environmental Sciences Undergraduate Theses

The main objective ofthis study was to determine the presence and level of mosaicism for chromosomal aneuploidy in several cases of fetal demise that had previous indications of chromosome aberrations. The presence of aneuploidy was determined using fluorescence in situ hybridization (FISH) probes. Techniques that were utilized in attempts to prepare several tissue types for FISH are described. The analyses made with FISH provided useful data on the chromosomal makeup oftwo of the cases studied. In one case FISH results showed that the product of conception (POC) did not contain the trisomy 18 that is present at low levels in ...


Development Of An Assay To Determine The Amount Of Binding Between Dna And The Carcinogen (+)-Anti-Benzo (A) Pyrene Diol Eopxide, Cher Wright Apr 1994

Development Of An Assay To Determine The Amount Of Binding Between Dna And The Carcinogen (+)-Anti-Benzo (A) Pyrene Diol Eopxide, Cher Wright

Life and Environmental Sciences Undergraduate Theses

A method for determining the number of carcinogenic molecules bound (adducted) to a DNA sequence was studied. Samples of purified plasmid DNA were exposed to increasing concentrations of the carcinogenic chemical benzo(a)pyrene diol epoxide (BPDE). Treated plasmids then served as the template in a Polymerase Chain Reaction (PCR). Desired test regions of the plasmid were selectively amplified. Samples from the PCR were then electrophoresed to study the number of successful copies produced by the PCR and the size and density of the bands. DNA replication levels decreased proportionately as BPDE exposure concentrations increased. The results suggest that the ...


Immunomagnetic Selection Of Acrosome-Reacted Bovine Spermatozoa, Thomas Mccormick Apr 1993

Immunomagnetic Selection Of Acrosome-Reacted Bovine Spermatozoa, Thomas Mccormick

Life and Environmental Sciences Undergraduate Theses

The anterior end of the mammalian sperm nucleus is covered by a thin, double-layered membranous sac that contains hydrolytic enzymes including acrosin, hyaluronidase, esterases and acid hydrolases. These enzymes, which are involved in the initial phase of fertilization, are activated and released during the acrosome reaction. This exposure of the acrosomal contents occurs not only during fertilization, but also during sperm death.

Acrosome-reacted sperm cells were selected from thawed, cryopreserved samples of bovine spermatozoa using rabbit antibodies to bovine acrosine and goat anti-rabbit immunoglobulin-coated magnetic beads. The selection process was monitored by staining with fluorescence isothiocynate- beads coupled to anti-rabbit ...


Enzyme Cytochemical Localization Of Na,K-Atpase And Immunohistochemical Localization Of Vinculin In Frog And Mouse Liver Tissue, Douglas Mcinnis Apr 1992

Enzyme Cytochemical Localization Of Na,K-Atpase And Immunohistochemical Localization Of Vinculin In Frog And Mouse Liver Tissue, Douglas Mcinnis

Life and Environmental Sciences Undergraduate Theses

Means of localizing molecules are essential in contemporary biological research attempting to unlock the mysteries of how cells function, interact, and go wrong. Enzyme cytochemistry and immunohistochemistry are two methods which utilize different characteristics of given molecules to localize them in tissue. The objective of this study was to localize two proteins, Na,K-ATPase and vinculin, in frog and mouse liver sections using these methods. Na,KATPase activity contributes to overall p-nitrophenylphosphatase (jpNPPase) activity, and sites of p-NPPase activity were determined using an enzyme cytochemical method. Sites of p-NPPase activity were thus extrapolated to indicate regions of probable Na,K-ATPase ...


Identification Of Laminin Domains That Promote Tumor Cell Adhesion, Corby Freitag Apr 1991

Identification Of Laminin Domains That Promote Tumor Cell Adhesion, Corby Freitag

Life and Environmental Sciences Undergraduate Theses

The process of metastasis requires tumor cells to migrate through the basement membrane which consists of laminin, a glycoprotein which promotes the adhesion of HT-1080 fibrosarcoma cells. The present study was designed to test four peptides, corresponding to short sequences of the intact laminin molecule, for cell adhesion-promoting activity. By using three different cell migration assays, the four peptides were monitored for their cell adhesive-promoting activity. Results were obtained showing some cell adhesive-promoting activity from all four peptides.


Bilirubin-Albumin Binding At Varying Conditions Of Hematocrit, Sodium Benzoate Concentration, And Bilirubin/Albumin Molar Ratios, Patrick Ahrens Apr 1990

Bilirubin-Albumin Binding At Varying Conditions Of Hematocrit, Sodium Benzoate Concentration, And Bilirubin/Albumin Molar Ratios, Patrick Ahrens

Life and Environmental Sciences Undergraduate Theses

Hyperbilirubinemia, which causes jaundice, is quite common in newborns. Typically bilirubin is bound to its carrier protein albumin and very little unbound bilirubin is present in the blood. However, in cases of hyperbilirubinemia there is a marked increase in unbound bilirubin, which can be toxic. The two methods traditionally used to treat hyperbilirubinemia are exchange transfusion and phototherapy. Each of these methods have drawbacks. The Department of Chemical, Bio, and Materials Engineering at Arizona State University is currently developing an extra-corporeal hemoperfusion column that will remove the unbound bilirubin fraction from the blood of jaundiced patients. This column utilizes coated ...


In Situ Hybridization Technique As Applied To Cytogenetics, Daniel Coverdell Apr 1988

In Situ Hybridization Technique As Applied To Cytogenetics, Daniel Coverdell

Life and Environmental Sciences Undergraduate Theses

In our experimentation with the in situ hybridization technique we were unable to band chromosomes with the standard Giemsa staining procedure following autoradiography. Banding was achieved using a replication banding technique in which bromodeoxyuridine (BrdU) was incorporated during replication. The background particles and debris on the chromosomal preparations, which had been in situ hybridized, was reduced by maintaining constant agitation during post-hybridization washes. Excessive background streaks, resulting from cracks in the radiography emulsion, were nearly eliminated by heating the emulsion to 50° C while dipping the slides.


A New Assay For 6-Pyruvoyl Tetrahydropterin Synthase From A Rat Kidney Model, John Connolly Jr. Apr 1988

A New Assay For 6-Pyruvoyl Tetrahydropterin Synthase From A Rat Kidney Model, John Connolly Jr.

Life and Environmental Sciences Undergraduate Theses

The enzyme 6-pyruvoyl tetrahydropterin synthase (PTP synthase) is the second enzyme in a four - step pathway which leads to the production of tetrahydrobiopterin, the cofactor for aromatic amino acid hydroxylations. In attempts to develop an assay, PTP synthase was characterized in terms of what effect its concentration, substrate (dihydroneopterin triphosphate-h^NTP) concentration, incubation time, and cofactor concentration had on product (sepiapterin) formation. In a reaction mixture of 100 pi, sepiapterin production is proportional to enzyme amount only up to 10 pi of enzyme. Sepiapterin production is linear to substrate concentration up to 60 pM after which, sepiapterin production levels off ...


In Situ Hybridization Of Complementary Deoxyribonucleic Acid Probes To The 1448-1455 Region Of Escherichia Coli 16s Ribosomal Subunits, Angela Lubbers Apr 1988

In Situ Hybridization Of Complementary Deoxyribonucleic Acid Probes To The 1448-1455 Region Of Escherichia Coli 16s Ribosomal Subunits, Angela Lubbers

Life and Environmental Sciences Undergraduate Theses

30S subunits were isolated from Escherichia coli ribosomes and purified 16S RNA was subsequently extracted. An eight-base DNA oligomer [d(CCCGAAGG)] was hybridized in situ to the 1448-1455 region of the 16S rRNA. The cDNA-rRNA hybrid was digested with the RNA specific enzyme RNase H. The clipped RNA fragments were then separated by polyacrylamide gel electrophoresis. The desired RNA band was visualized by UV shadowing, eluted from the gel and sequenced to verify the specificity of the probe binding. The results suggest that the cDNA and rRNA were binding, but proof that it was specific to the 1448-1455 region is ...