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Full-Text Articles in Molecular Biology

Safety And Potency Testing Of Veterinary Vaccines , Louise Mary Henderson Jan 1993

Safety And Potency Testing Of Veterinary Vaccines , Louise Mary Henderson

Retrospective Theses and Dissertations

Prior to licensing of veterinary vaccines, it is essential to demonstrate that the use of the product is safe (e.g., use of the product will not have unexpected or undesirable effects on animals exposed to the agent or on the environment) and potent (e.g., produces the desired result). The projects described in this dissertation demonstrate research directed toward the development and improvement of safety and potency testing of veterinary vaccines from a regulatory perspective;In the first study, recombination between vaccine strains of pseudorabies virus (PRV) resulting in the creation of strains with undesirable virulence and antigenic combinations ...


Genetic And Biochemical Characterization Of The P21[Superscript Ras]-Related Protein Rho1p In The Yeast Saccharomyces Cerevisiae , Joni Marie Seeling Johnson Jan 1992

Genetic And Biochemical Characterization Of The P21[Superscript Ras]-Related Protein Rho1p In The Yeast Saccharomyces Cerevisiae , Joni Marie Seeling Johnson

Retrospective Theses and Dissertations

Rho1p is a guanine nucleotide binding protein coded for by the Saccharomyces cerevisiae gene RHO1, which is homologous to p21[superscript] ras and other members of the ras superfamily of small molecular weight GTP binding proteins. This protein family is highly conserved in evolution and in some instances is known to be involved in control of cell proliferation. The precise physiological functions of most p21[superscript] ras-related proteins, however, are not known. In order to gain further insight into the physiological function of rho proteins, this research addressed the genetic and biochemical characterization of Rho1p, as well as the subcellular ...


Cloning And Characterization Of Mink Plasminogen Activator Inhibitor Type 1 (Pai-1) Cdna And The Regulation Of Mink Pai-1 Expression At Mrna Level , Tsung-Hsien Chuang Jan 1992

Cloning And Characterization Of Mink Plasminogen Activator Inhibitor Type 1 (Pai-1) Cdna And The Regulation Of Mink Pai-1 Expression At Mrna Level , Tsung-Hsien Chuang

Retrospective Theses and Dissertations

Mink plasminogen activator inhibitor type 1 (PAI-1) is a glycoprotein with molecular weight about 50 kD and is highly regulated in cultured cells. In mink lung CCL64 epithelial cells, epidermal growth factor (EGF), transforming growth factor-[beta] (TGF-[beta]), and 12-O-tetradecanoyl phorbol-13-acetate (TPA) increase the synthesis of PAI-1. Epidermal growth factor and TGF-[beta] synergistically stimulate the synthesis of PAI-1. The synergistic effect and factors which govern the PAI-1 expression in mink lung CCL64 cells were studied by using a mink PAI-1 cDNA as a probe to detect PAI-1 mRNA;The mink PAI-1 cDNA was cloned from a cDNA library ...


Characterization Of The Saccharomyces Cerevisiae Gene Mgp1, A Novel Member Of The Ras Gene Superfamily That Functions In The Mitochondrial Biogenesis , Shu-Hwa Lee Tzeng Jan 1992

Characterization Of The Saccharomyces Cerevisiae Gene Mgp1, A Novel Member Of The Ras Gene Superfamily That Functions In The Mitochondrial Biogenesis , Shu-Hwa Lee Tzeng

Retrospective Theses and Dissertations

This dissertation characterizes Mgp1p, a mitochondrial protein that functions in mitochondrial biogenesis. Mgp1p is the first protein related to the p21[superscript] H-ras oncoprotein shown to function in mitochondria;The Saccharomyces cerevisiae gene MGP1 was isolated based on its ability to restore respiratory competence to a strain containing the nuclear petite mutation msg1-1. MGP1 codes for a protein of approximately 58 kDa, with an amino terminal sequence typical of a mitochondrial targeting peptide. A 190 residue region of Mgp1p shares approximately 30% amino acid sequence identity with any member of p21[superscript] ras-related protein superfamily, with particularly strong homology in ...


Studies On The Role Of Expression Of Phosphoenolpyruvate Carboxykinase Gene In The Development Of Bovine Lactation Ketosis , Sowmya Moorthi Jan 1992

Studies On The Role Of Expression Of Phosphoenolpyruvate Carboxykinase Gene In The Development Of Bovine Lactation Ketosis , Sowmya Moorthi

Retrospective Theses and Dissertations

Changes in expression of the phosphoenolpyruvate carboxykinase (PEPCK) gene, which codes for a key regulatory enzyme of gluconeogenesis, were studied in bovine liver tissue. Induction of ketosis was attempted in cows by a protocol of restricting feed intake (FR) and supplementing with 1,3-butanediol (BD), a ketogenic precursor, beginning at 15 d postpartum (FRBD protocol). Twenty Holstein cows were distributed in equal numbers among four treatments: normal control, obese control, normal cows given FRBD, and obese cows given FRBD. Liver biopsies were obtained at nine different times during early lactation: 10 d prepartum, 5 and 14 d postpartum (just before ...


Analysis Of Three Plasmids From Filamentous, Nitrogen-Fixing Cyanobacteria , Douglas Kevin Walton Jan 1991

Analysis Of Three Plasmids From Filamentous, Nitrogen-Fixing Cyanobacteria , Douglas Kevin Walton

Retrospective Theses and Dissertations

The cyanobacteria are an ancient and diverse group of prokaryotes capable of oxygenic photosynthesis. They display both unicellular and filamentous cell types and some have the ability to fix atmospheric dinitrogen, and the ability to differentiate specialized cells. In addition, many of the cyanobacteria possess plasmids. To date, only one plasmid, from a unicellular non-nitrogen-fixer, has been characterized in detail. The current lack of information concerning cyanobacterial plasmids, and their subsequent use as vectors, has hindered genetic analysis, particularly in the filamentous species;In this study, the properties of two plasmids and one plasmid origin of replication from filamentous, nitrogen-fixing ...


Effect Of 1,10-Phenanthroline On Dna Binding Of 1,25-Dihydroxyvitamin D Receptors And Characterization And Regulation Of Retinoic Acid Receptors In Rat Osteosarcoma Cells , Kevin Brady Atkins Jan 1991

Effect Of 1,10-Phenanthroline On Dna Binding Of 1,25-Dihydroxyvitamin D Receptors And Characterization And Regulation Of Retinoic Acid Receptors In Rat Osteosarcoma Cells , Kevin Brady Atkins

Retrospective Theses and Dissertations

The effect of the chelator 1,10-phenanthroline on the DNA-binding of 1,25-dihydroxyvitamin D receptor was demonstrated. Increasing concentrations of the chelator, up to 3 mM, decreased receptor binding to DNA-cellulose. This effect was specific for the chelating function because the nonchelating analog, 1,7-phenanthroline, did not inhibit DNA-cellulose binding of the receptor. The cations, Cd[superscript]2+ and Zn[superscript]2+, were able to block chelator-mediated inhibition. These cations were unable to reverse the inhibition unless 1,10-phenanthroline was removed by dialysis;The rat osteosarcoma 17/2.8 cells were shown to express both the cellular retinoic acid binding ...


Avena Phytochrome Mrna: In Vivo Localization, Degradation Products, And Half-Life , Kevin Andrew Seeley Jan 1991

Avena Phytochrome Mrna: In Vivo Localization, Degradation Products, And Half-Life , Kevin Andrew Seeley

Retrospective Theses and Dissertations

The purpose of this research is to determine the role that post-transcriptional processes play in the regulation of phytochrome (phy) steady-state messenger RNA (mRNA) levels. The control of etiolated phytochrome (phyA) and chlorophyll a/b binding protein (cab) gene transcription and steady-state mRNA levels are known to be regulated by the phyA signal transduction pathway. I performed a number of control experiments that indicate the degradation products detected in northern analysis of phyA mRNA were produced in vivo and were not an artifact of RNA isolation techniques. I have also shown that infrared (IR) irradiation has no appreciable effect on ...


Identification Of The Immediate-Early Products Of Bovine Herpesvirus-1 , David A. Hulce Jan 1990

Identification Of The Immediate-Early Products Of Bovine Herpesvirus-1 , David A. Hulce

Retrospective Theses and Dissertations

Bovine herpesvirus-1 (BHV-1) is a member of the alphaherpesvirinae. Productive infections of the alphaherpesvirinae progress through three stages of viral transcription and translation. The first stage, immediate-early (IE), does not require prior viral gene expression and is required for the second, delayed-early (DE), and third, late (L), stages of the infectious cycle. Like other herpesviruses, BHV-1 frequently establishes latent infections. This study was undertaken to characterize the IE stage of the productive BHV-1 infection. The number and location of IE genes and the size of IE polypeptides and IE mRNAs were investigated;Probing genomic Southern blots of BHV-1 cloned in ...


Regulation Of Protein Expression By Transforming Growth Factor Beta And Other Growth Modulators , Frederic William Thalacker Jan 1990

Regulation Of Protein Expression By Transforming Growth Factor Beta And Other Growth Modulators , Frederic William Thalacker

Retrospective Theses and Dissertations

The main goal of this project was to understand the signal transduction pathways through which growth modulators such as TGF-[beta], PMA and retinoic acid mediate protein expression and DNA synthesis. Transforming growth factor [beta] (TGF-[beta]) induces synthesis and secretion of a 48,000 M r protein that we have found to be type I plasminogen activator inhibitor (PAI-1). TGF-[beta] induces PAI-1 in every cell line that we have studied including near term mink lung epithelial CCL64 cells, mouse embryo fibroblast AKR-2B 84A cells, African green monkey kidney epithelial BSC-1 cells, and normal rat kidney fibroblast NRK 49F ...


Genetic And Molecular Analyses Of Suppressor Of Forked In Drosophila Melanogaster , Cynthia Lynn Fitch-Steenson Jan 1990

Genetic And Molecular Analyses Of Suppressor Of Forked In Drosophila Melanogaster , Cynthia Lynn Fitch-Steenson

Retrospective Theses and Dissertations

Mutations at the suppressor of forked, su(f), locus are trans-acting, allele specific suppressors and enhancers of mutations at certain other loci. Su(f) is a trans-acting regulator whose normal product is necessary for proper transcription of these genes. The su(f) product regulates the transcription of transposable elements located within these genes. Our investigation includes a genetic analysis in which a number of new su(f) alleles were isolated, a development analysis in which the effects of new alleles and allelic combinations were characterized, and a molecular analysis in which a cloned genomic DNA fragment containing su(f) was ...


Isolation Of A Chicken Cardiac Muscle Titin Cdna Probe , Greg Robert Sater Jan 1990

Isolation Of A Chicken Cardiac Muscle Titin Cdna Probe , Greg Robert Sater

Retrospective Theses and Dissertations

The objective of this project was to construct and identify a cDNA probe for the striated muscle protein titin. RNA was isolated from skeletal and cardiac muscle of 16 day chicken embryos using a guanidinium isothiocyanate procedure. The RNA thus isolated was both intact (as shown by agarose gel electrophoresis) and transcribable in vitro; additionally, it was possible to obtain very large RNA molecules in this manner. cDNA was then constructed utilizing random oligodeoxynucleotides for priming of the first strand synthesis reaction, instead of the more common oligo-dT priming. Random priming was also utilized for second strand synthesis. As with ...


The Metabolism And Localization Of Two Proteins During Heat Shock Of Tetrahymena Thermophila , Thomas William Mcmullin Jan 1987

The Metabolism And Localization Of Two Proteins During Heat Shock Of Tetrahymena Thermophila , Thomas William Mcmullin

Retrospective Theses and Dissertations

When T. thermophila is subjected to hyperthermal stress, it immediately but transiently represses the synthesis of the normal array of proteins (non-hsps) and induces the synthesis of the heat shock protein (hsps). These changes in protein synthetic patterns are controlled at both the transcriptional and translational levels;To determine whether the translational regulatory events could be correlated with changes in the protein synthetic machinery, heat shock induced modifications of the ribosomal proteins were investigated. Of particular interest was the heat shock induced association of a 22 kDa protein, p22, with the small subunit of the ribosome. The synthesis of p22 ...


The Mouse Major Histocompatibility Complex And Age-Related Changes In The Murine Immune System , Terry Euclaire Meyer Jan 1987

The Mouse Major Histocompatibility Complex And Age-Related Changes In The Murine Immune System , Terry Euclaire Meyer

Retrospective Theses and Dissertations

A and C57BL/6 inbred mice were used as models for investigating interactions between aging processes and the genes that regulate the immune system, the major histocompatibility complex (MHC, H-2) genes. The C57BL/6 (H-2[superscript] b) mice exhibited significantly longer mean lifespans than the A (H-2[superscript] a) mice, as has been observed by other investigators. Backcross and F[subscript] 2 generation offspring from the parental A and C57BL/6 mice were tested for dependence of longevity upon H-2 type. The effects of H-2 type upon lifespan were statistically significant only for the oldest surviving (i.e., "10[superscript ...


The Heat Shock Response In Tetrahymena Thermophila: Control Of Protein Synthesis , Kevin W. Kraus Jan 1986

The Heat Shock Response In Tetrahymena Thermophila: Control Of Protein Synthesis , Kevin W. Kraus

Retrospective Theses and Dissertations

The changes in protein synthesis which result when Tetrahymena thermophila cells are shifted from 30(DEGREES)C to 40(DEGREES)C are shown to be regulated at both the mRNA accumulation and mRNA translation levels. The heat shock protein mRNAs are preferentially translated early in a heat shock while non-heat shock protein messages remain in the cell in a non-translated state. Later in heat shock these non-hsp messages return to polysomes at the same time as a small heat-induced RNA becomes associated with polysomal ribosomes. This novel RNA species may play an important role in the return to normal protein ...


Production And Use Of Monoclonal Antibodies For Molecular Characterization, Affinity Purification, And Serological Detection Of Economically Significant Plant Viruses , Robert Diaco Jan 1985

Production And Use Of Monoclonal Antibodies For Molecular Characterization, Affinity Purification, And Serological Detection Of Economically Significant Plant Viruses , Robert Diaco

Retrospective Theses and Dissertations

Hybridomas secreting monoclonal antibodies (M-Abs) against three isolates of barley yellow dwarf virus (BYDV) were established. Two M-Abs were generated against the MAV isolate, one against RPV, and six against P-PAV. None of the M-Abs reacted with healthy host components. Reactions of M-Abs, or unlabelled polyclonal antisera, in an indirect enzyme-linked immunosorbent assay (ELISA) indicate the presence of a common epitope on all three virus isolates. BYDV dissociated when incubated in carbonate-bicarbonate coating buffer at pH 9.6; stabilization was achieved by predialysis with either 2% formaldehyde or 2% glutaraldehyde. In indirect ELISA, unlabelled polyclonal antisera bound to both stabilized ...