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Molecular Biology

1995

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Articles 1 - 30 of 49

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

Molecular Characterization Of Mitofilin, A Novel, Mitochondrial, Coiled Coil Protein, And The Relationship Between Organism Complexity And Coiled Coil Protein-Mediated Structure: A Dissertation, Paul R. Odgren Nov 1995

Molecular Characterization Of Mitofilin, A Novel, Mitochondrial, Coiled Coil Protein, And The Relationship Between Organism Complexity And Coiled Coil Protein-Mediated Structure: A Dissertation, Paul R. Odgren

GSBS Dissertations and Theses

In the course of experiments designed to identify and characterize structural proteins of the nuclear matrix, one antibody was generated which recognized an extraction-resistant cytoplasmic protein. This antibody was used as the starting point in the cloning and molecular characterization of a novel protein of the inter-membrane space of the mitochondrion which has been named mitofilin. Mitofilin is expressed in all human cell types, and murine homologues also exist. Mitofilin associates only with mitochondria and not with other membrane-bounded organelles such as Golgi or endoplasmic reticulum. This observation has been confirmed both by biochemical fractionation and multi-label fluorescence microscopy. Recombinant ...


Anaerobic Protoporphyrin Biosynthesis Does Not Require Incorporation Of Methyl Groups From Methionine, David Bollivar Oct 1995

Anaerobic Protoporphyrin Biosynthesis Does Not Require Incorporation Of Methyl Groups From Methionine, David Bollivar

Scholarship

No abstract provided.


The Essential Ost2 Gene Encodes The 16-Kd Subunit Of The Yeast Oligosaccharyltransferase, A Highly Conserved Protein Expressed In Diverse Eukaryotic Organisms, Susana Silberstein, Pallia G. Collins, Daniel J. Kelleher, Reid Gilmore Oct 1995

The Essential Ost2 Gene Encodes The 16-Kd Subunit Of The Yeast Oligosaccharyltransferase, A Highly Conserved Protein Expressed In Diverse Eukaryotic Organisms, Susana Silberstein, Pallia G. Collins, Daniel J. Kelleher, Reid Gilmore

Open Access Articles

Oligosaccharyltransferase catalyzes the transfer of a preassembled high mannose oligosaccharide from a dolichol-oligosaccharide donor to consensus glycosylation acceptor sites in newly synthesized proteins in the lumen of the rough endoplasmic reticulum. The Saccharomyces cerevisiae oligosaccharyltransferase is an oligomeric complex composed of six non-identical subunits (alpha-zeta). The alpha, beta, gamma, and delta subunits of the oligosaccharyltransferase are encoded by the OST1, WBP1, OST3, and SWP1 genes, respectively. Here we describe the functional characterization of the OST2 gene that encodes the epsilon-subunit of the oligosaccharyltransferase. Genomic disruption of the OST2 locus was lethal in haploid yeast showing that expression of the Ost2 ...


Biochemical And Molecular Characterization Of The Prostate-Specific Membrane Antigen (Psma), John Karl Troyer Oct 1995

Biochemical And Molecular Characterization Of The Prostate-Specific Membrane Antigen (Psma), John Karl Troyer

Theses and Dissertations in Biomedical Sciences

Prostate cancer is the most common malignancy and the second leading cause of cancer death in males in the United States. Additionally, the number of deaths attributed to prostate cancer is increasing at a rate of approximately 8% a year. Development of new diagnostic and therapy strategies are needed in order to improve the life expectancy of patients with this disease. One tool which may allow for improvements in prostate cancer diagnosis and therapy is the monoclonal antibody (MAb) 7E11-C5.3 which was first described in 1987. Since then, the antigen recognized by MAb 7E11-C5.3 has been named the ...


Anaerobic Protoporphyrin Biosynthesis Does Not Require Incorporation Of Methyl Groups From Methionine, David Bollivar, Thomas Elliott, Samuel I. Beale Sep 1995

Anaerobic Protoporphyrin Biosynthesis Does Not Require Incorporation Of Methyl Groups From Methionine, David Bollivar, Thomas Elliott, Samuel I. Beale

David Bollivar

It was recently reported (H. Akutsu, J.-S. Park, and S. Sano, J. Am. Chem. Soc. 115:12185–12186, 1993) that in the strict anaerobe methyl groups from exogenous L-methionine are incorporated specifically into the 1 and 3 positions (Fischer numbering system) on the heme groups of cytochrome c3. It was suggested that under anaerobic conditions, protoporphyrin IX biosynthesis proceeds via a novel pathway that does not involve coproporphyrinogen III as a precursor but instead may use precorrin-2 (1,3-dimethyluroporphyrinogen III), a siroheme and vitamin B12 precursor which is known to be derived from uroporphyrinogen III via methyl transfer from ...


The Cytoplasmic Tail Domain Of The Vacuolar Protein Sorting Receptor Vps10p And A Subset Of Vps Gene Products Regulate Receptor Stability, Function, And Localization, Joan Lin-Cereghino, Eric G. Marcusson, Scott D. Emr Sep 1995

The Cytoplasmic Tail Domain Of The Vacuolar Protein Sorting Receptor Vps10p And A Subset Of Vps Gene Products Regulate Receptor Stability, Function, And Localization, Joan Lin-Cereghino, Eric G. Marcusson, Scott D. Emr

College of the Pacific Faculty Articles

VPS10 of Saccharomyces cerevisiae encodes a type I transmembrane receptor protein required for the sorting of the soluble vacuolar hydrolase carboxypeptidase Y (CPY). To characterize the essential structural features and intercompartmental transport itinerary of the CPY receptor, we have constructed mutant forms of Vps10p that alter the carboxyterminal cytoplasmic tail of the protein. In addition, we have analyzed the effect these mutations as well as mutations in several VPS genes have on the function, stability, and localization of Vps10p. Although wild-type Vps10p is very stable over a 3-h chase period, overproduction of Vps10p results in PEP4-dependent degradation of the receptor ...


Light-Dependent And Tissue-Specific Expression Of The H-Protein Of The Glycine Decarboxylase Complex, R. Srinivasan, David J. Oliver Aug 1995

Light-Dependent And Tissue-Specific Expression Of The H-Protein Of The Glycine Decarboxylase Complex, R. Srinivasan, David J. Oliver

David J. Oliver

Glycine decarboxylase is a mitochondrial enzyme complex, which is the site of photorespiratory CO2 and NH3 release. Although the proteins that constitute the complex are located within the mitochondria, because of their intimate association with photosynthesis their expression is controlled by light. Comparisons of the kinetics of mRNA accumulation between the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and the H-protein of glycine decarboxylase during the greening of etiolated Arabidopsis thaliana suggest that their expression is controlled in parallel. A genomic clone for the H-protein (gdcH) was isolated from Arabidopsis and sequenced. The upstream region from -856 to +62 was ...


Functional Characterization Of Ost3p. Loss Of The 34-Kd Subunit Of The Saccharomyces Cerevisiae Oligosaccharyltransferase Results In Biased Underglycosylation Of Acceptor Substrates, Denise Karaoglu, Daniel J. Kelleher, Reid Gilmore Aug 1995

Functional Characterization Of Ost3p. Loss Of The 34-Kd Subunit Of The Saccharomyces Cerevisiae Oligosaccharyltransferase Results In Biased Underglycosylation Of Acceptor Substrates, Denise Karaoglu, Daniel J. Kelleher, Reid Gilmore

Open Access Articles

Within the lumen of the rough endoplasmic reticulum, oligosaccharyltransferase catalyzes the en bloc transfer of a high mannose oligosaccharide moiety from the lipid-linked oligosaccharide donor to asparagine acceptor sites in nascent polypeptides. The Saccharomyces cerevisiae oligosaccharyltransferase was purified as a heteroligomeric complex consisting of six subunits (alpha-zeta) having apparent molecular masses of 64 kD (Ost1p), 45 kD (Wbp1p), 34 kD, 30 kD (Swp1p), 16 kD, and 9 kD. Here we report a structural and functional characterization of Ost3p which corresponds to the 34-kD gamma-subunit of the oligosaccharyltransferase. Unlike Ost1p, Wbp1p, and Swp1p, expression of Ost3p is not essential for viability ...


Investigation Of The Substrate Recognition Characteristics And Kinetics Of Mammalian Mitochondrial Dna Topoisomerase I, Zeki Topcu Jul 1995

Investigation Of The Substrate Recognition Characteristics And Kinetics Of Mammalian Mitochondrial Dna Topoisomerase I, Zeki Topcu

Theses and Dissertations in Biomedical Sciences

Topoisomerases are DNA-modifying enzymes found in prokaryotes, eukaryotes, viruses and organelles such as chloroplast and mitochondria. Information about these enzymes in eukaryotic systems is mostly limited to nuclear enzymes, although our laboratory has been characterizing the biochemical and biophysical properties of the mammalian mitochondrial topoisomerases. We have determined the polarity of the attachment of mitochondrial topoisomerase I to its substrate DNA. To study the substrate preference and kinetic parameters of mitochondrial topoisomerase I, selected regions of mammalian mitochondrial DNA (mtDNA) were inserted into pGEM plasmid vectors following a series of modification and optimization experiments of currently available methods for PCR-cloning ...


Identification And Characterization Of Mitochondrial Dna Variants In Alzheimer's Disease, Natasha Singh Hamblet Jul 1995

Identification And Characterization Of Mitochondrial Dna Variants In Alzheimer's Disease, Natasha Singh Hamblet

Theses and Dissertations in Biomedical Sciences

Alzheimer's Disease (AD) is a complex neurodegenerative disorder that affects a significant portion of the human population regardless of ethnicity or gender. A mitochondrial hypothesis of AD has been proposed based on a number of studies which establish altered oxidative phosphorylation (OXPHOS) and ATP synthesis in AD tissue. ATP demand is most prevalent in the brain; damage to OXPHOS could severely impair brain metabolism, thereby leading to a decline in cognitive function. Four out of five complexes in the OXPHOS pathway are partly encoded by mitochondrial DNA (mtDNA); thus, this may be a crucial site of lesions that alter ...


In Vitro Expression Of Human Zona Pellucida Protein 3, Chih-Wei Chen Jul 1995

In Vitro Expression Of Human Zona Pellucida Protein 3, Chih-Wei Chen

Theses and Dissertations in Biomedical Sciences

Human zona pellucida protein 3 (hZP3) is the putative receptor on the zona pellucida of the mature oocyte that recognizes and binds sperm, and therefore plays a critical role in fertilization.

A full length cDNA of hZP3 (1278 bp) was amplified from the human ovary mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The hZP3 cDNA was subcloned into PSK and pREP4 expression vectors. The cDNA of hZP3 was further characterized by restriction mapping, PCR, auto-sequencing and Southern blot analysis by using an internal oligonucleotide probe, and found to be identical to the one reported by J. Dean. Using autosequencing, 289 ...


Moraxella (Branhamella) Catarrhalis: A Molecular Epidemiology Study, Lyndell R. Gill May 1995

Moraxella (Branhamella) Catarrhalis: A Molecular Epidemiology Study, Lyndell R. Gill

Electronic Theses and Dissertations

Moraxella (Branhamella) catarrhalis is the third-most-frequently isolated microorganism associated with acute exacerbations of chronic bronchitis in patients during their stay at the Mountain Home VA Medical Center (MHVAMC). In order to develop a practical, epidemiologically-meaningful typing method for M. (B.) catarrhalis, we tested two methods based on analysis of chromosomal DNA for typeability, reproducibility, and ability to differentiate between unrelated strains (discriminatory power, D). M. (B.) catarrhalis isolants from MHVAMC from 7/1/87-6/30/88 were grown overnight in broth and embedded in agarose. DNA was isolated by standard methods. The DNA was subjected to: (1) restriction endonuclease digestion ...


An "Extended -10" Promoter Alone Directs The Transcription Of The Dpnii Operon Of Streptococcus Pneumoniae, Alexander G. Sabelnikov, Bill Greenberg, Sanford Lacks Apr 1995

An "Extended -10" Promoter Alone Directs The Transcription Of The Dpnii Operon Of Streptococcus Pneumoniae, Alexander G. Sabelnikov, Bill Greenberg, Sanford Lacks

Alexander G Sabelnikov

No abstract provided.


Pathogenicity Of Murine Cytomegalovirus Mutants, Victoria Jean Cavanaugh Apr 1995

Pathogenicity Of Murine Cytomegalovirus Mutants, Victoria Jean Cavanaugh

Theses and Dissertations in Biomedical Sciences

The purpose of this study was to identified nonessential murine cytomegalovirus (MCMV) genes involved in pathogenesis in vivo. Our approach to identifyjng these genes consisted of constructing MCMV mutants, and then analyzing these mutants in vitro and in vivo. Recombinant viruses (RV) expressing the β-glucuronidase marker gene were constructed by site-directed insertion and deletion mutagenesis of the MCMV Hind III-J and -I regions of the viral genome. Mutations were targeted to this region of the MCMV genome because the corresponding region of the human CMV genome is nonessential and is involved in down-regulating major histocompatibility complex (MHC) class I expression ...


Identification And Characterization Of Genes Associated With V-Jun Induced Cell Transformation, Martin Toralballa Hadman Apr 1995

Identification And Characterization Of Genes Associated With V-Jun Induced Cell Transformation, Martin Toralballa Hadman

Biological Sciences Theses & Dissertations

The v-jun oncogene was initially identified as the causative agent for fibrosarcomas in chickens. Studies show that overexpression of v-Jun proteins transforms chicken embryo fibroblasts (CEF) in vitro, and forms tumors in chickens in vivo. The mechanisms for this are not clearly defined. Conceivably, overexpression of an unregulated transcription factor would cause cell transfonnation by illicit regulation of its target genes. In support of this, we show that in vivo v-Jun complexes exhibit differential binding to in vitro generated AP-1 and 'AP-1 like' target sequences, suggesting that the pattern of target gene expression is altered during cell transformation. With this ...


Sensitization To Self (Virus) Antigen By In Situ Expression Of Murine Interferon-Gamma., Myung-Shik Lee, Matthias Von Herrath, Hans Reiser, Michael B.A. Oldstone, Nora Sarvetnick Feb 1995

Sensitization To Self (Virus) Antigen By In Situ Expression Of Murine Interferon-Gamma., Myung-Shik Lee, Matthias Von Herrath, Hans Reiser, Michael B.A. Oldstone, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Autoimmune disease results from inflammatory destruction of tissues by aberrant self-reactive lymphocytes. We studied the autoimmune potential of T lymphocytes immunologically ignorant of viral antigens acting as self antigens and whether the host defense molecule IFN-gamma could stimulate these cells to cytotoxic competency. For this purpose, we produced double transgenic mice expressing pancreatic IFN-gamma as well as lymphocytic choriomeningitis virus (LCMV) nucleoprotein (NP) or glycoprotein (GP) antigen. 100% of the NP+/IFN-gamma+ mice became diabetic before 2 mo of age, while none of the NP single transgenic littermates and only 10% of IFN-gamma single transgenic littermates did. Strikingly, NP+/IFN-gamma ...


Myasthenia Gravis-Like Syndrome Induced By Expression Of Interferon Gamma In The Neuromuscular Junction., Danling Gu, Lise Wogensen, Nigel A. Calcutt, Chunyao Xia, Simin Zhu, John P. Merlie, Howard S. Fox, Jon Lindstrom, Henry C. Powell, Nora Sarvetnick Feb 1995

Myasthenia Gravis-Like Syndrome Induced By Expression Of Interferon Gamma In The Neuromuscular Junction., Danling Gu, Lise Wogensen, Nigel A. Calcutt, Chunyao Xia, Simin Zhu, John P. Merlie, Howard S. Fox, Jon Lindstrom, Henry C. Powell, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Abnormal humoral responses toward motor end plate constituents in muscle induce myasthenia gravis (MG). To study the etiology of this disease, and whether it could be induced by host defense molecules, we examined the consequences of interferon (IFN) gamma production within the neuromuscular junction of transgenic mice. The transgenic mice exhibited gradually increasing muscular weakness, flaccid paralysis, and functional disruption of the neuromuscular junction that was reversed after administration of an inhibitor of acetylcholinesterase, features which are strikingly similar to human MG. Furthermore, histological examination revealed infiltration of mononuclear cells and autoantibody deposition at motor end plates. Immunoprecipitation analysis indicated ...


The Alpha Subunit Of The Saccharomyces Cerevisiae Oligosaccharyltransferase Complex Is Essential For Vegetative Growth Of Yeast And Is Homologous To Mammalian Ribophorin I, Susana Silberstein, Pallia G. Collins, Daniel J. Kelleher, Peter J. Rapiejko, Reid Gilmore Feb 1995

The Alpha Subunit Of The Saccharomyces Cerevisiae Oligosaccharyltransferase Complex Is Essential For Vegetative Growth Of Yeast And Is Homologous To Mammalian Ribophorin I, Susana Silberstein, Pallia G. Collins, Daniel J. Kelleher, Peter J. Rapiejko, Reid Gilmore

Open Access Articles

Oligosaccharyltransferase mediates the transfer of a preassembled high mannose oligosaccharide from a lipid-linked oligosaccharide donor to consensus glycosylation acceptor sites in newly synthesized proteins in the lumen of the rough endoplasmic reticulum. The Saccharomyces cerevisiae oligosaccharyltransferase is an oligomeric complex composed of six nonidentical subunits (alpha-zeta), two of which are glycoproteins (alpha and beta). The beta and delta subunits of the oligosaccharyltransferase are encoded by the WBP1 and SWP1 genes. Here we describe the functional characterization of the OST1 gene that encodes the alpha subunit of the oligosaccharyltransferase. Protein sequence analysis revealed a significant sequence identity between the Saccharomyces cerevisiae ...


Formation Of The Isocyclic Ring Of Chlorophyll By Isolated Clamydomonas Reinhardtii Chloroplasts, David Bollivar, Samuel I. Beale Jan 1995

Formation Of The Isocyclic Ring Of Chlorophyll By Isolated Clamydomonas Reinhardtii Chloroplasts, David Bollivar, Samuel I. Beale

David Bollivar

Chlamydomonas reinhardtii chloroplasts catalyzed two sequential steps of Chl biosynthesis, S-adenosyl-L-methionine:Mg-protoporphyrin IX methyltransferase and Mg-protoporphyrin IX monomethyl ester oxidative cyclase. A double mutant strain of C. reinhardtii was constructed which has a cell wall deficiency and is unable to form chlorophyll in the dark. Dark-grown cells were disrupted with a BioNeb nebulizer under conditions which lysed the plasma membrane but not the chloroplast envelope. Chloroplasts were purified by Percoll density gradient centrifugation. The purified chloroplasts were used to define components required for the biosynthesis of Mg-2,4-divinylpheoporphyrin a 5 (divinyl protochlorophyllide) from Mg-protoporphyrin IX. Product formation requires the addition ...


A Small Modified Hammerhead Ribozyme And Its Conformational Characteristics Determined By Mutagenesis And Lattice Calculation, Brooke Lustig, N H. Lin, S M. Smith, R L. Jernigan, K.-T Jeang Jan 1995

A Small Modified Hammerhead Ribozyme And Its Conformational Characteristics Determined By Mutagenesis And Lattice Calculation, Brooke Lustig, N H. Lin, S M. Smith, R L. Jernigan, K.-T Jeang

Brooke S. Lustig

A prototypic hammerhead ribozyme has three helices that surround an asymmetrical central core loop. We have mutagenized a hammerhead type ribozyme. In agreement with previous studies, progressive removal of stem-loop II from a three stemmed ribozyme showed that this region is not absolutely critical for catalysis. However, complete elimination of stem II and its loop did reduce, but did not eliminate, function. In a stem-loop II-deleted ribozyme, activity was best preserved when a purine, preferably a G, was present at position 10.1. This G contributed to catalysis irregardless of its role as either one part of a canonical pair ...


Consistencies Of Individual Dna Base-Amino Acid Interactions In Structures And Sequences, Brooke Lustig, R L. Jernigan Jan 1995

Consistencies Of Individual Dna Base-Amino Acid Interactions In Structures And Sequences, Brooke Lustig, R L. Jernigan

Brooke S. Lustig

Amino acid-amino acid interaction energies have been derived from crystal structure data for a number of years. Here is reported the first derivation of normalized relative interaction from binding data for each of the four bases interacting with a specific amino acid, utilizing data from combinatorial multiplex ONA binding of zinc finger domains [Desjarlais, J. R. and Berg, J. M. (1994) Proc. Natl. Acad. Sci. USA, 91, 11099–11103]. The five strongest Interactions are observed for lysine-guanine, lysine-thymine, arginine- guanine, aspartic acid-cytosine and asparagineadenine. These rankings for interactions with the four bases appear to be related to base-amino acid partial ...


Formation Of The Isocyclic Ring Of Chlorophyll By Isolated Clamydomonas Reinhardtii Chloroplasts, David Bollivar Jan 1995

Formation Of The Isocyclic Ring Of Chlorophyll By Isolated Clamydomonas Reinhardtii Chloroplasts, David Bollivar

Scholarship

No abstract provided.


Consistencies Of Individual Dna Base-Amino Acid Interactions In Structures And Sequences, Brooke Lustig, R L. Jernigan Jan 1995

Consistencies Of Individual Dna Base-Amino Acid Interactions In Structures And Sequences, Brooke Lustig, R L. Jernigan

Faculty Publications, Chemistry

Amino acid-amino acid interaction energies have been derived from crystal structure data for a number of years. Here is reported the first derivation of normalized relative interaction from binding data for each of the four bases interacting with a specific amino acid, utilizing data from combinatorial multiplex ONA binding of zinc finger domains [Desjarlais, J. R. and Berg, J. M. (1994) Proc. Natl. Acad. Sci. USA, 91, 11099–11103]. The five strongest Interactions are observed for lysine-guanine, lysine-thymine, arginine- guanine, aspartic acid-cytosine and asparagineadenine. These rankings for interactions with the four bases appear to be related to base-amino acid partial ...


A Small Modified Hammerhead Ribozyme And Its Conformational Characteristics Determined By Mutagenesis And Lattice Calculation, Brooke Lustig, N H. Lin, S M. Smith, R L. Jernigan, K.-T Jeang Jan 1995

A Small Modified Hammerhead Ribozyme And Its Conformational Characteristics Determined By Mutagenesis And Lattice Calculation, Brooke Lustig, N H. Lin, S M. Smith, R L. Jernigan, K.-T Jeang

Faculty Publications, Chemistry

A prototypic hammerhead ribozyme has three helices that surround an asymmetrical central core loop. We have mutagenized a hammerhead type ribozyme. In agreement with previous studies, progressive removal of stem-loop II from a three stemmed ribozyme showed that this region is not absolutely critical for catalysis. However, complete elimination of stem II and its loop did reduce, but did not eliminate, function. In a stem-loop II-deleted ribozyme, activity was best preserved when a purine, preferably a G, was present at position 10.1. This G contributed to catalysis irregardless of its role as either one part of a canonical pair ...


Genetic And Biochemical Characterization Of Brucella Abortus Strain Rb51 , Allen Elroy Jensen Jan 1995

Genetic And Biochemical Characterization Of Brucella Abortus Strain Rb51 , Allen Elroy Jensen

Retrospective Theses and Dissertations

Brucella abortus strain RB51, a naturally derived rough variant, has been proposed to replace strain 19 for use as a vaccine against brucellosis in cattle. Utilizing fingerprinting and standard biotyping techniques, it was possible to differentiate RB51 from other brucellae and to ascertain the stability of RB51. Standardization of a genomic fingerprinting procedure using restriction endonuclease profiles of total DNA from Brucella are described. Although the focus was principally on B. abortus isolates from cattle, the investigation included field isolates from bison and elk, reference strains, vaccinal strains, and genetically-engineered brucellae;Pulsed-field gel electrophoresis profiles of the reference type strains ...


Characterization Of A Rat Clonal [Beta]-Cell Line, Rinm5f, In A Passage- And Morphology-Dependent Manner , Bum-Sup Lee Jan 1995

Characterization Of A Rat Clonal [Beta]-Cell Line, Rinm5f, In A Passage- And Morphology-Dependent Manner , Bum-Sup Lee

Retrospective Theses and Dissertations

A novel insulin-like immunoreactivity, adrenergic receptors (ARs) and voltage-dependent Ca[superscript]2+ channels (VDCCs) were characterized in a clonal [beta]-cell line, RINm5F, in a passage- and morphology-dependent manner. The cells exhibited two morphological features; the differentiated cells (DCs) with long neuron-like processes and the undifferentiated cells (UCs) without the processes;As morphology of the cells changed from DCs to UCs, amount of secreted insulin gradually decreased; however, insulin reacted to antiserum vs. bovine insulin (ABI) gradually increased from barely detectable amounts. Immunoblotting and immunocytochemistry showed that antiserum vs. rat insulin reacted with insulin from both cell types. ABI did ...


Identification And Characterization Of G4-Dna Binding Proteins From Tetrahymena Thermophila , Ted Patrick Schierer Jan 1995

Identification And Characterization Of G4-Dna Binding Proteins From Tetrahymena Thermophila , Ted Patrick Schierer

Retrospective Theses and Dissertations

Alternative DNA conformations have been known to exist since before the elucidation of the double helix. The study of alternative DNA conformations formed by telomeric sequences has led to detailed studies of biologically relevant DNA capable of forming unusual structures. In order to understand the biological role of alternative DNA conformations formed by telomeric DNA, proteins which bind these structures have been identified and characterized;The initial portion of my research involved the identification of a protein from Tetrahymena thermophila which binds preferentially to parallel-stranded G-quartet structures. Competition and binding assays demonstrated that the protein, TGP (Tetrahymena G4 binding protein ...


Characterization And Application Of A Novel Cholesterol-Reducing Anaerobe, Eubacterium Coprostanoligenes Atcc 51222 , Ling Li Jan 1995

Characterization And Application Of A Novel Cholesterol-Reducing Anaerobe, Eubacterium Coprostanoligenes Atcc 51222 , Ling Li

Retrospective Theses and Dissertations

A novel cholesterol-reducing bacterium, Eubacterium coprostanoligenes ATCC 51222, was characterized and tested for its hypocholesterolemic effects in animal models. The bacterium did not require cholesterol for growth and synthesized cholesterol reductase constitutively. Lecithin was required for growth but excess lecithin in media did not improve bacterial growth and coprostanol production. Addition of pyruvate, lactose, and most reducing agents in media increased bacterial growth and coprostanol production. The bacterium was sensitive to most antibiotics but had strong resistance to tetracycline. A resting cell assay was established to evaluate the cholesterol reductase activity. E. coprostanoligenes reduced cholesterol through an indirect pathway. Intermediates ...


Expression And Characterization Of Bovine Rotavirus Proteins In The Baculovirus System , Young Soo Lyoo Jan 1995

Expression And Characterization Of Bovine Rotavirus Proteins In The Baculovirus System , Young Soo Lyoo

Retrospective Theses and Dissertations

Recombinant proteins of bovine rotavirus VP2, VP4, VP6 and VP7 were expressed in insect cells using baculovirus expression vectors. Expressed recombinant proteins were similar to those of rotavirus. Recombinant viral antigens expressed in insect cells reacted with polyclonal antiserum to the VMRI strain of BRV by an indirect immunofluorescence test. Expressed recombinant proteins were characterized by radioimmunoprecipitation and their molecular mass was found to be similar to that of in wild type rotavirus. These four recombinant proteins were antigenic in guinea pigs and induced antibodies to the corresponding rotavirus proteins as detected by indirect immunofluorescence test. Recombinant viruses Bac.pVL ...


Calsensin, A Novel Calcium-Binding Protein Expressed In A Subset Of Peripheral Leech Neurons Fasciculating In A Single Axon Tract , Kristen Kay Briggs Jan 1995

Calsensin, A Novel Calcium-Binding Protein Expressed In A Subset Of Peripheral Leech Neurons Fasciculating In A Single Axon Tract , Kristen Kay Briggs

Retrospective Theses and Dissertations

The lan 3-6 monoclonal antibody recognizes a subpopulation of peripheral neurons whose afferents segregate into a single fascicle within the developing leech central nervous system. The lan 3-6 monoclonal antibody was used to isolate a clone encoding a novel protein, calsensin, which contains two EF-hand calcium-binding domains. In situ hybridizations with calsensin riboprobes, Western blots of lan 3-6 monoclonal antibody immunoprecipitations, and immunohistochemistry with polyclonal antibodies generated against a partially-purified calsensin fusion protein confirmed that the sequence encoding the lan 3-6 antigen has been identified. Partially-purified calsensin fusion protein binds [superscript]45Ca[superscript]2+ in vitro, suggesting that the calsensin ...