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Molecular Biology

1997

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Articles 1 - 30 of 53

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

Activation Of A Retroviral Membrane Fusion Protein: Soluble Receptor-Induced Liposome Binding Of The Alsv Envelope Glycoprotein, Lorraine D. Hernandez, Reuben J. Peters, Sue E. Delos, John A. T. Young, David A. Agard, Judith M. White Dec 1997

Activation Of A Retroviral Membrane Fusion Protein: Soluble Receptor-Induced Liposome Binding Of The Alsv Envelope Glycoprotein, Lorraine D. Hernandez, Reuben J. Peters, Sue E. Delos, John A. T. Young, David A. Agard, Judith M. White

Reuben J. Peters

It is not known how membrane fusion proteins that function at neutral pH, for example the human immunodeficiency virus envelope (Env) glycoprotein and intracellular fusion machines, are activated for target bilayer binding. We have addressed this question using a soluble oligomeric form of an avian retroviral Env glycoprotein (API) and soluble forms of its receptor. Binding of soluble receptor to API induces API to bind to liposomes composed of phosphatidylcholine and cholesterol at neutral pH. Liposome binding only occurs at fusion permissive temperatures (T > 20°C), is complete between 2 to 5 min at 37°C, and is stable to ...


Sex Chromosome Ratios And Aneuploidy Rates In The Zona Pellucida Selected Spermatozoa From Normal And Subfertile Men, Qinuo Fan Van Dyk Oct 1997

Sex Chromosome Ratios And Aneuploidy Rates In The Zona Pellucida Selected Spermatozoa From Normal And Subfertile Men, Qinuo Fan Van Dyk

Theses and Dissertations in Biomedical Sciences

The human hemizona has been demonstrated to select spermatozoa with good motility, normal morphology, and the capacity to undergo the zona-induced acrosomal reaction. The studies conducted here are directed at using the human hemizona to investigate two key questions: (1) whether human X-chromosome bearing spermatozoa (X-sperm) and Y-chromosome bearing spermatozoa (Y-sperm) differ in their functional survival time (assessed by their capacity to bind to the human zona pellucida) after prolonged in vitro incubation, and (2) whether hemizona-bound spermatozoa have a reduced aneuploidy rate compared to unbound (and therefore, unselected) spermatozoa?

In the functional survival study, donor spermatozoa were held in ...


Collaborative Roles For C-Jun N-Terminal Kinase, C-Jun, Serum Response Factor, And Sp1 In Calcium-Regulated Myocardial Gene Expression, Patrick M. Mcdonough, Deanna S. Hanford, Amy B. Sprenkle, Noel R. Mellon, Christopher C. Glembotski Sep 1997

Collaborative Roles For C-Jun N-Terminal Kinase, C-Jun, Serum Response Factor, And Sp1 In Calcium-Regulated Myocardial Gene Expression, Patrick M. Mcdonough, Deanna S. Hanford, Amy B. Sprenkle, Noel R. Mellon, Christopher C. Glembotski

Amy Sprenkle

Electrical stimulation of contractions (pacing) of primary neonatal rat ventricular myocytes increases intracellular calcium and activates a hypertrophic growth program that includes expression of the cardiac-specific gene, atrial natriuretic factor (ANF). To investigate the mechanism whereby pacing increases ANF, pacing was tested for its ability to regulate mitogen-activated protein kinase family members, ANF promoter activity, and the trans-activation domain of the transcription factor, Sp1. Pacing and the calcium channel agonist BAYK 8644 activated c-Jun N-terminal kinase (JNK) but not extracellular signal-regulated kinase. Pacing stimulated ANF-promoter activity approximately 10-fold. Furthermore, transfection with an expression vector for c-Jun, a substrate for JNK ...


Interferon Gamma (Ifn-Gamma) Is Necessary For The Genesis Of Acetylcholine Receptor-Induced Clinical Experimental Autoimmune Myasthenia Gravis In Mice., Balaji Balasa, Caishu Deng, Jae Lee, Linda M. Bradley, Dyanna K. Dalton, Premkumar Christadoss, Nora Sarvetnick Aug 1997

Interferon Gamma (Ifn-Gamma) Is Necessary For The Genesis Of Acetylcholine Receptor-Induced Clinical Experimental Autoimmune Myasthenia Gravis In Mice., Balaji Balasa, Caishu Deng, Jae Lee, Linda M. Bradley, Dyanna K. Dalton, Premkumar Christadoss, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Experimental autoimmune myasthenia gravis (EAMG) is an animal model of human myasthenia gravis (MG). In mice, EAMG is induced by immunization with Torpedo californica acetylcholine receptor (AChR) in complete Freund's adjuvant (CFA). However, the role of cytokines in the pathogenesis of EAMG is not clear. Because EAMG is an antibody-mediated disease, it is of the prevailing notion that Th2 but not Th1 cytokines play a role in the pathogenesis of this disease. To test the hypothesis that the Th1 cytokine, interferon (IFN)-gamma, plays a role in the development of EAMG, we immunized IFN-gamma knockout (IFN-gko) (-/-) mice and wild-type ...


Ms2 Coat Protein Mutants Which Bind Qβ Rna, Marc Spingola, David S. Peabody Jul 1997

Ms2 Coat Protein Mutants Which Bind Qβ Rna, Marc Spingola, David S. Peabody

Marc Spingola

The coat proteins of the RNA phages MS2 and Qβ are structurally homologous, yet they specifically bind different RNA structures. In an effort to identify the basis of RNA binding specificity we sought to isolate mutants that convert MS2 coat protein to the RNA binding specificity of Qβ. A library of mutations was created which selectively substitutes amino acids within the RNA binding site. Genetic selection for the ability to repress translation from the Qβ translational operator led to the isolation of several MS2 mutants that acquired binding activity for Qβ RNA. Some of these also had reduced abilities to ...


Differential Glycosylation Of Tractin And Leechcam, Two Novel Ig Superfamily Members, Regulates Neurite Extension And Fascicle Formation, Yueqiao Huang, John Jellies, Kristen M. Johansen, Jorgen Johansen Jul 1997

Differential Glycosylation Of Tractin And Leechcam, Two Novel Ig Superfamily Members, Regulates Neurite Extension And Fascicle Formation, Yueqiao Huang, John Jellies, Kristen M. Johansen, Jorgen Johansen

Zoology and Genetics Publications

By immunoaffinity purification with the mAb Lan3-2, we have identified two novel Ig superfamily members, Tractin and LeechCAM. LeechCAM is an NCAM/FasII/ApCAM homologue, whereas Tractin is a cleaved protein with several unique features that include a PG/YG repeat domain that may be part of or interact with the extracellular matrix. Tractin and LeechCAM are widely expressed neural proteins that are differentially glycosylated in sets and subsets of peripheral sensory neurons that form specific fascicles in the central nervous system. In vivo antibody perturbation of the Lan3-2 glycoepitope demonstrates that it can selectively regulate extension of neurites and ...


The Cellular And Molecular Dynamics Of The Queuosine Modification In Transfer Rna: Definition, Modulation, Deficiencies And Effect Of The Queuosine Modification System, Rana C. Morris Jul 1997

The Cellular And Molecular Dynamics Of The Queuosine Modification In Transfer Rna: Definition, Modulation, Deficiencies And Effect Of The Queuosine Modification System, Rana C. Morris

Theses and Dissertations in Biomedical Sciences

The presence of the queuosine modification in the wobble position of tRNAasn, tRNasp, tRNAhis, and tRNAtyr is associated with a decrease in cellular growth rate, an increase in the ability to withstand environmental stress, and differentiation of pleuripotent cells into mature phenotypes. The loss of this normal modification is strongly correlated with neoplastic transformation and tumor progression of a wide variety of cancers.

The "normal" system for formation of the queuosine modification in tRNA was studied in human fibroblast cell cultures and in mouse, rat and human liver tissues. The queuosine modification system is shown to ...


Interaction Of Acth And Estradiol In The Regulation Of Growth, Differentiation, And Steroidogenic Maturation Of The Baboon Fetal Adrenal Gland, Maria Gomez Leavitt Jul 1997

Interaction Of Acth And Estradiol In The Regulation Of Growth, Differentiation, And Steroidogenic Maturation Of The Baboon Fetal Adrenal Gland, Maria Gomez Leavitt

Theses and Dissertations in Biomedical Sciences

The present study determined whether the growth of, as well as ACTH receptor and P-450 enzyme messenger ribonucleic acid and/or protein levels in the baboon fetal adrenal are dependent upon fetal pituitary ACTH during mid and late gestation and the mechanism by which placental estrogen modulates ACTH actions. Administration of betamethasone (3mg/day) to baboon mothers on days 60-99 of gestation and to the fetus (0.6 mg/, n = 4) or to the fetus (0.6mg) and mother (6 mg/ml; n = 4) every other day between days 150-164 of gestation (term = 184 days) decreased (P $53β-hydroxysteroid dehydrogenase-isomerase (3β-HSD ...


Type Ii Estrogen Binding-Site Agonist: The Synthesis And Biological Evaluation Of The Enantiomers Of Methyl-Para-Hydroxyphenyllactate (Mehpla), Lester G. Pretlow Jul 1997

Type Ii Estrogen Binding-Site Agonist: The Synthesis And Biological Evaluation Of The Enantiomers Of Methyl-Para-Hydroxyphenyllactate (Mehpla), Lester G. Pretlow

Theses and Dissertations in Biomedical Sciences

The function of the type II estrogen binding site (EBS) has yet to be determined. However, a high affinity ligand for the binding site has been identified through HPLC and GC-MS. This ligand, MeHPLA, bears a structural relationship to a group of compounds called "phytoestrogens" which, along with MeHPLA, have been observed to suppress the cellular proliferation of estrogen sensitive MCF-7 breast cancer cells in vitro. Additionally, MeHPLA has been observed to suppress the growth of rat uteri in vivo. The high affinity of MeHPLA for the type II EBS suggests that this interaction is responsible for the observed suppression ...


Fluorescence In Situ Hybridization Analysis Of Human Embryos Derived From In Vitro And In Vivo Matured Oocytes, Constance Descisciolo Jul 1997

Fluorescence In Situ Hybridization Analysis Of Human Embryos Derived From In Vitro And In Vivo Matured Oocytes, Constance Descisciolo

Theses and Dissertations in Biomedical Sciences

Despite adequate hormonal stimulation, oocytes collected for the purpose of in vitro fertilization and embryo transfer display several levels of nuclear maturity. Preovulatory or mature oocytes, technically those that are Metaphase I or II, are inseminated shortly after aspiration and assessed for fertilization the following day. Prophase I oocytes, also called germinal vesicle-bearing or immature oocytes, require a 24-36 hour period in culture before being exposed to spermatozoa. During this time, the majority of Prophase I oocytes complete nuclear maturation in vitro, progressing from germinal vesicle breakdown through first polar body extrusion. If inseminated, many in vitro matured oocytes fertilize ...


Oral Cancer Screening, Damon Omar Watson Jun 1997

Oral Cancer Screening, Damon Omar Watson

Honors Theses

Cancers of the oral cavity and pharynx have increased throughout the decades. Since oral cancer represents a small percentage of the total cancers in the United States, it is often overlooked. Though this is the case, the results of this cancer can be devastating. To address this issue, an Oral Cancer Screening was organized on August 14, 1996 at the West Michigan Cancer Center in Kalamazoo. This was a joint collaboration between the author, the West Michigan Cancer Center, and the Kalamazoo Valley Dental Society. This event allowed research to take place concerning the data obtained. Confidential results and information ...


Molecular Basis Of The Mechanism And Regulation Of Receptor-Gtp Binding Protein Interactions: A Thesis, Marianne Wessling-Resnick Jun 1997

Molecular Basis Of The Mechanism And Regulation Of Receptor-Gtp Binding Protein Interactions: A Thesis, Marianne Wessling-Resnick

GSBS Dissertations and Theses

The photon receptor, rhodopsin, and the GTP-binding regulatory protein, transducin, belong to a family of G protein-coupled receptors. The activation process through which guanine nucleotide exchange of the G protein is accomplished was investigated utilizing these components of the visual transduction system. Rhodopsin, modelled as an enzyme in its interaction with substrates, transducin and guanine nucleotides, was characterized to catalyze the G protein's activation by a double-displacement mechanism. Remarkable allosteric behavior was observed in these kinetic studies. Equilibrium binding studies were performed to investigate the molecular basis of the positive cooperative behavior between transducin and rhodopsin. These experiments show ...


Local Delivery Of Interleukin 4 By Retrovirus-Transduced T Lymphocytes Ameliorates Experimental Autoimmune Encephalomyelitis., Michael K. Shaw, James B. Lorens, Archana Dhawan, Richard Dalcanto, Harley Y. Tse, Alyssa B. Tran, Colleen Bonpane, Shanti L. Eswaran, Stefan Brocke, Nora Sarvetnick, Lawrence Steinman, Garry P. Nolan, C. Garrison Fathman May 1997

Local Delivery Of Interleukin 4 By Retrovirus-Transduced T Lymphocytes Ameliorates Experimental Autoimmune Encephalomyelitis., Michael K. Shaw, James B. Lorens, Archana Dhawan, Richard Dalcanto, Harley Y. Tse, Alyssa B. Tran, Colleen Bonpane, Shanti L. Eswaran, Stefan Brocke, Nora Sarvetnick, Lawrence Steinman, Garry P. Nolan, C. Garrison Fathman

Journal Articles: Regenerative Medicine

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune disease of the central nervous system which serves as a model for the human disease multiple sclerosis. We demonstrate here that encephalitogenic T cells, transduced with a retroviral gene, construct to express interleukin 4, and can delay the onset and reduce the severity of EAE when adoptively transferred to myelin basic protein-immunized mice. Thus, T lymphocytes transduced with retroviral vectors can deliver "regulatory cytokines" in a site-specific manner and may represent a viable therapeutic strategy for the treatment of autoimmune disease.


Search For Water-Soluble, Nontoxic Fullerene Derivatives To Inhibit Hiv-I Protease Activity, Christopher J. Leichliter May 1997

Search For Water-Soluble, Nontoxic Fullerene Derivatives To Inhibit Hiv-I Protease Activity, Christopher J. Leichliter

All-College Writing Contest

No abstract provided.


Endogenous Alkylglycerol Functions As A Mediator Of Protein Kinase C Activity And Cell Proliferation, Fritz G. Buchanan May 1997

Endogenous Alkylglycerol Functions As A Mediator Of Protein Kinase C Activity And Cell Proliferation, Fritz G. Buchanan

Electronic Theses and Dissertations

To explore the possibility that 1-O-alkyl-sn-glycerol (alkylglycerol) may serve a regulatory role in the control of cell proliferation or PKC activity, we examined the ability of alkylglycerol to influence PKC activity and subcellular distribution as well as the ability of alkylglycerol to effect cell proliferation. MDCK cells grown to confluence show a loss of PKC activity associated with the membrane, as reported in fibroblasts. Preconfluent cultures of MDCK cells have a high level of PKC activity associated with the membrane. However, treatment of preconfluent cultures with alkylglycerol causes a reduction of PKC activity. A similar inhibition was observed with alkylglycerol ...


Endosome To Golgi Retrieval Of The Vacuolar Protein Sorting Receptor, Vps10p, Requires The Function Of The Vps29, Vps30, And Vps35 Gene Products, Matthew N. J. Seaman, Eric G. Marcusson, Joan Lin-Cereghino, Scott D. Emr Apr 1997

Endosome To Golgi Retrieval Of The Vacuolar Protein Sorting Receptor, Vps10p, Requires The Function Of The Vps29, Vps30, And Vps35 Gene Products, Matthew N. J. Seaman, Eric G. Marcusson, Joan Lin-Cereghino, Scott D. Emr

College of the Pacific Faculty Articles

Mutations in the S. cerevisiae VPS29 and VPS30 genes lead to a selective protein sorting defect in which the vacuolar protein carboxypeptidase Y (CPY) is missorted and secreted from the cell, while other soluble vacuolar hydrolases like proteinase A (PrA) are delivered to the vacuole. This phenotype is similar to that seen in cells with mutations in the previously characterized VPS10 and VPS35 genes. Vps10p is a late Golgi transmembrane protein that acts as the sorting receptor for soluble vacuolar hydrolases like CPY and PrA, while Vps35p is a peripheral membrane protein which cofractionates with membranes enriched in Vps10p. The ...


Cγ- And Cα-Subunits Provide The Specificity For Camp/Pka Function, Wei Qing Zhang Apr 1997

Cγ- And Cα-Subunits Provide The Specificity For Camp/Pka Function, Wei Qing Zhang

Theses and Dissertations in Biomedical Sciences

The Cα and Cγ of the cAMP-dependent protein kinase (PKA) are two highly homologous (83% amino acid identity), yet functionally distinct isozymes in vitro for substrate and pseudosubstrate specificity, and in intact cells for cell phenotypes (Beebe, 1992).

To determine the molecular mechanisms underlying the distinct functions in cell phenotypes, in vitro experiments were designed to make a detailed comparison of Cγ and Cα for substrate and pseudosubstrate specificity. To this end, Cγ and Cα were expressed m mammalian cells, bacteria and Sf9 insect cells using baculovirus. Abundant expression of active enzyme was cell system specific. Cγ and Cα expressed ...


Ifn-Gamma, Igif, And Iddm., Nora Sarvetnick Feb 1997

Ifn-Gamma, Igif, And Iddm., Nora Sarvetnick

Journal Articles: Regenerative Medicine

No abstract provided.


Introduction Of A [4fe-4s (S-Cys)4]+1,+2 Iron-Sulfur Center Into A Four-Α Helix Protein Using Design Parameters From The Domain Of The Fx Cluster In The Photosystem I Reaction Center, Marvin Paul Scott, John Biggins Feb 1997

Introduction Of A [4fe-4s (S-Cys)4]+1,+2 Iron-Sulfur Center Into A Four-Α Helix Protein Using Design Parameters From The Domain Of The Fx Cluster In The Photosystem I Reaction Center, Marvin Paul Scott, John Biggins

Agronomy Publications

We describe the insertion of an iron-sulfur center into a designed four α-helix model protein. The model protein was re-engineered by introducing four cysteine ligands required for the coordination of the mulinucleate cluster into positions in the main-chain directly analogous to the domain predicted to ligand the interpeptide [4Fe-4S (S-cys)4] cluster, Fx, from PsaA and PsaB of the Photosystem I reaction center. This was achieved by inserting the sequence, CDGPGRGGTC, which is conserved in PsaA and PsaB, into interhelical loops 1 and 3 of the four α-helix model. The holoprotein was characterized spectroscopically after insertion of the iron-sulfur center ...


Rna Base-Amino Acid Interaction Strengths Derived From Structures And Sequences, Brooke Lustig, S Arora, R L. Jernigan Jan 1997

Rna Base-Amino Acid Interaction Strengths Derived From Structures And Sequences, Brooke Lustig, S Arora, R L. Jernigan

Brooke S. Lustig

We investigate RNA base-amino acid interactions by counting their contacts in structures and their implicit contacts in various functional sequences where the structures can be assumed to be preserved. These frequencies are cast into equations to extract relative interaction energetics. Previously we used this approach in considering the major groove interactions of DNA, and here we apply it to the more diverse interactions observed in RNA. Structures considered are the three different tRNA synthetase complexes, the U1A spliceosomal protein with an RNA hairpin and the BIV TARTat complex. We use binding data for the base frequencies for the seryl, aspartyl ...


Rna Base-Amino Acid Interaction Strengths Derived From Structures And Sequences, Brooke Lustig, S Arora, R L. Jernigan Jan 1997

Rna Base-Amino Acid Interaction Strengths Derived From Structures And Sequences, Brooke Lustig, S Arora, R L. Jernigan

Faculty Publications, Chemistry

We investigate RNA base-amino acid interactions by counting their contacts in structures and their implicit contacts in various functional sequences where the structures can be assumed to be preserved. These frequencies are cast into equations to extract relative interaction energetics. Previously we used this approach in considering the major groove interactions of DNA, and here we apply it to the more diverse interactions observed in RNA. Structures considered are the three different tRNA synthetase complexes, the U1A spliceosomal protein with an RNA hairpin and the BIV TARTat complex. We use binding data for the base frequencies for the seryl, aspartyl ...


Gene Expression Studies Of Murine Pregnancy Associated Granulated Metrial Gland And Decidual Cells , Melissa Phillips Allen Jan 1997

Gene Expression Studies Of Murine Pregnancy Associated Granulated Metrial Gland And Decidual Cells , Melissa Phillips Allen

Retrospective Theses and Dissertations

During pregnancy in viviparous mammals, the maternal decidua develops between the embryo and the uterine endometrium. This dissertation describes research on two different cell types present within the rodent decidua during pregnancy, namely, granulated metrial gland (GMG) cells and decidual cells;GMG cells are natural killer cells that proliferate and differentiate within the murine uterus during pregnancy. Their differentiation is associated with the accumulation of cytolytic mediators within cytoplasmic granules. The signaling mechanisms required for GMG cell differentiation, however, are largely unknown. In this study, expression of the genes coding for granzymes D, E, F, and G (granzymes D-G) was ...


The Drosophila Melanogaster Rad54 Homolog, Dmrad54, Is Involved In The Repair Of Radiation Damage And Recombination, Rolf Kooistra, José B. M. Zonneveld, Anja De Jong, Jan C. J. Eeken, Chris J. Osgood, Jean-Marie Buerstedde, Paul H. M. Lohman, Albert Pastink Jan 1997

The Drosophila Melanogaster Rad54 Homolog, Dmrad54, Is Involved In The Repair Of Radiation Damage And Recombination, Rolf Kooistra, José B. M. Zonneveld, Anja De Jong, Jan C. J. Eeken, Chris J. Osgood, Jean-Marie Buerstedde, Paul H. M. Lohman, Albert Pastink

Biological Sciences Faculty Publications

The RAD54 gene of Saccharomyces cerevisiae plays a crucial role in recombinational repair of double-strand breaks in DNA. Here the isolation and functional characterization of the RAD54 homolog of the fruit fly Drosophila melanogaster, DmRAD54, are described. The putative Dmrad54 protein displays 46 to 57% identity to its homologs from yeast and mammals. DmRAD54 RNA was detected at all stages of fly development, but an increased level was observed in early embryos and ovarian tissue. To determine the function of DmRAD54, a null mutant was isolated by random mutagenesis. DmRAD54-deficient flies develop normally, but the females are sterile. Early ...


Protein Engineering Of The Ph Dependence And Substrate Specificity Of Glucoamylase From Aspergillus Awamori , Tsuei-Yun Fang Jan 1997

Protein Engineering Of The Ph Dependence And Substrate Specificity Of Glucoamylase From Aspergillus Awamori , Tsuei-Yun Fang

Retrospective Theses and Dissertations

Glucoamylase (EC 3.2.1.3) (GA), an enzyme that catalyzes the release of [beta]-D-glucose from the nonreducing ends of starch and related oligo- and polysaccharides, is widely used in industry to produce high-glucose syrups. Site-directed mutagenesis was used to construct mutations to obtain mutant enzymes with industrially desired alterations in pH dependence and substrate specificity;An increased pH optimum for GA would be industrially desirable so that it could be used in process conditions more similar to that of other enzymatic steps in starch processing. Five mutations were designed to remove or weaken the hydrogen bond between Ser411 ...


Host Regulation Of The Replication Of The Yeast Retrotransposon Ty5 , Ning Ke Jan 1997

Host Regulation Of The Replication Of The Yeast Retrotransposon Ty5 , Ning Ke

Retrospective Theses and Dissertations

The replication of retroelements may have deleterious effects on the hosts. Therefore, elements and their hosts have coevolved mechanisms to regulate retroelement replication. This regulation can occur at the transcriptional, translational, or post-translational levels as well as through targeted integration or cDNA recombination. I have studied host regulation of the Saccharomyces retrotransposon Ty5. I have found that Ty5 transcription is carefully controlled. Transcription of Ty5 is haploid specific and is repressed 10 fold in diploid strains. Ty5 transcription is also regulated by the pheromone response pathway, which is activated during mating. In addition, Ty5 transcription is regulated by flanking sequences ...


Genetic Analysis Of Chlorophyll Biosynthesis, David Bollivar Jan 1997

Genetic Analysis Of Chlorophyll Biosynthesis, David Bollivar

Scholarship

No abstract provided.


Cloning, Expression And Bioactivity Of Porcine Soluble Tnf Receptor 1 And The Comparison Of Pk(15) And Wehi 164(13)-Based Tnf Bioassays , Nancy A. Maroushek Boury Jan 1997

Cloning, Expression And Bioactivity Of Porcine Soluble Tnf Receptor 1 And The Comparison Of Pk(15) And Wehi 164(13)-Based Tnf Bioassays , Nancy A. Maroushek Boury

Retrospective Theses and Dissertations

Tumor necrosis factor (TNF) is a pro-inflammatory cytokine that activates both leukocytes and endothelium and facilitates the movement of inflammatory cells from circulation to sites of infection or injury. This activation may be beneficial and aid in host defense, or may be detrimental and mediate part of the pathophysiology of disease. Several porcine diseases, such as salmonellosis and mycoplasmal pneumonia, cause an increase in TNF production, which may be either beneficial or harmful to swine health. Two related studies were completed to develop assays and reagents to further study the role of TNF in porcine disease;In the first study ...


Organization And Expression Of The Tup Gene In Daucus Carota , Philip Earl Dykema Jan 1997

Organization And Expression Of The Tup Gene In Daucus Carota , Philip Earl Dykema

Retrospective Theses and Dissertations

Differential screening of a somatic embryo cDNA library for genes expressed during embryogenesis has disclosed a small RNA which I have named Tiny Up-regulated Protein (TUP). This mRNA is 402 bp in length, is expressed during embryo formation, and reaches its highest accumulation in the flowers and roots of adult carrot plants. The mRNA is barely detectable in leaves. The accumulation of the TUP mRNA in seedlings is increased by auxin and by abscisic acid. The open reading frame encodes a tiny, uniquely structured protein of 6100 Daltons which has a pI of 5.3. Algorithms predicting protein structure suggest ...


Structure-Function Study Of A Plant Protein Proteinase Inhibitor: Site-Directed Mutagenesis Of Cucurbita Maxima Trypsin Inhibitor V , R. Sandra Benavente Jan 1997

Structure-Function Study Of A Plant Protein Proteinase Inhibitor: Site-Directed Mutagenesis Of Cucurbita Maxima Trypsin Inhibitor V , R. Sandra Benavente

Retrospective Theses and Dissertations

Cucurbita marima Trypsin Inhibitor-V (CMTI-V), isolated from pumpkin, is an inhibitor of trypsin and [beta]-Factor XIIa (FXIIa). CMTI-V binds to enzyme as would a substrate, but whereas substrate is cleaved at its scissile bond and the resulting fragments released, CMTI-V remains bound to its target enzyme, thus inhibiting activity. Amino acids at the amino side of the scissile bond are referred to as P1, P2, etc., away from the bond. Carboxyl-side amino acids are referred to as P1', P2', etc., away from the bond. We focused on the inhibitor amino acid positions P1' (D45) and P3 (V42). CMTI-V belongs ...


Use Of Pyrimidine Pathway To Produce Mutant Plants That Fail To Respond To Wound Induction , Lan Zhou Jan 1997

Use Of Pyrimidine Pathway To Produce Mutant Plants That Fail To Respond To Wound Induction , Lan Zhou

Retrospective Theses and Dissertations

To understand how proteinase inhibitor genes, as a part of plant defense system, are activated following wounding, we have developed a molecular genetic approach to directly select for mutants blocked in wound response in Arabidopsis thaliana. The wound-inducible promoter from the potato Proteinase Inhibitor 11 gene (pin2) was linked to a bacterial negative marker gene codA encoding for cytosine deaminase and transformed into Arabidopsis thaliana. The cytosine deaminase activity was detected in the leaves of the transgenic plants induced by sucrose, ABA, methyl jasmonate and mechanical wounding. Methyl jasmonate was the best for the pin2 induction in the seedlings among ...