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Full-Text Articles in Molecular Biology

Investigation Of The Molecular Mechanism Of Fertility Restoration In Texas Cytoplasm Maize , David Stewart Skibbe Jan 2004

Investigation Of The Molecular Mechanism Of Fertility Restoration In Texas Cytoplasm Maize , David Stewart Skibbe

Retrospective Theses and Dissertations

Cytoplasmic male sterility (cms) is a maternally inherited inability to produce viable pollen. In Texas (T)-cytoplasm maize, the combined action of two restorer of fertility genes, rfl and rfla , can overcome the male sterile phenotype. Previous research established rf2a encodes a mitochondrial aldehyde dehydrogenase (ALDH) with broad substrate specificity. This broad substrate specificity complicates determining the molecular mechanism of rf2a-mediated fertility restoration. In response, a combination of transgenic, biochemical, reverse genetic, and genomic approaches were utilized. Three additional functional maize ALDH genes (rf2b, rf2c, and rf2d) with differing substrate specificities were identified. Each ALDH gene was characterized via in ...


Studying The Replication Mechanism Of The Yeast Retrotransposon Ty5 By Molecular And Computational Approaches, Xiang Gao Jan 2001

Studying The Replication Mechanism Of The Yeast Retrotransposon Ty5 By Molecular And Computational Approaches, Xiang Gao

Retrospective Theses and Dissertations

The yeast retrotransposon Ty5 is a Ty1/copia element. It is in the Hemivirus genus of the Pseudoviridae family. The ability to genetically manipulate retrotransposons and the yeast host cell was taken advantage of to explore replication mechanisms unique to Ty5 and common to most retrotransposons. Because of the abundance and diversity of retroelement sequences, along with the fact that many retroelement enzymes have evolved unique functional specificities, computational approaches were also developed to study functional divergence in replication. By screening a randomly mutagenized Ty5 library, two mutations (Y68C, D252N) causing higher transposition frequencies were identified. Both mutations increased Ty5 ...


Genetic Mobility And Instability Of Retroviral Vector In Vector Producer Cells For Gene Therapy, Won-Bin Young Jan 1999

Genetic Mobility And Instability Of Retroviral Vector In Vector Producer Cells For Gene Therapy, Won-Bin Young

Retrospective Theses and Dissertations

A primary biosafety issue of retroviral vector-mediated gene therapy is the genetic instability of retroviral vectors. Reverse transcription of vector RNA genome is initiated by viral reverse transcriptase (RT) in a virion particle after infection of a target cell. During reverse transcription, abnormal template switches between vector and occasionally co-packaged helper virus in a virion particle can therefore enable helper virus to regain replication elements from the vector and revert to replication-competent retrovirus (RCR). This research was undertaken to study the origins of RT enzyme activities and test the hypothesis that RT enzyme activities are contributed by both exogenous RT ...


Investigating The Mechanism Of Mrp/Plf Gene Expression By Bfgf , Alesia Marie Hruska-Hageman Jan 1998

Investigating The Mechanism Of Mrp/Plf Gene Expression By Bfgf , Alesia Marie Hruska-Hageman

Retrospective Theses and Dissertations

The mitogen-regulated protein/proliferin (mrp/ plf) genes belong to the prolactin/growth hormone gene superfamily and encode at least four closely related proteins. Identified functions of these proteins include stimulation of uterine proliferation and endothelial angiogenesis. In 3T3 cells, basic fibroblast growth factor (bFGF) stimulates the production of mrp/plf mRNAs with a resulting increase in the protein products. Although the three cloned mrp/ plf gene promoters (mrp3, plf42, and plf149 ) are over 97% identical in sequence, only mrp3 is transcriptionally activated by bFGF. We have identified a sequence in the mrp3 promoter, which we have named the "bFGF-responsive element ...


Genetic And Biochemical Analysis Of Aspergillus Awamori Glucoamylase Thermostability , Martin John Allen Jan 1997

Genetic And Biochemical Analysis Of Aspergillus Awamori Glucoamylase Thermostability , Martin John Allen

Retrospective Theses and Dissertations

Aspergillus awamori glucoamylase (GA) is an enzyme involved in industrial corn starch processing. Genetic and biochemical approaches were used to study the mechanisms governing GA thermostability. Three proline substitution (Xaa → Pro) mutations were constructed that were predicted to increase the enzyme's stability by decreasing its conformational entropy of unfolding. When expressed in Saccharomyces cerevisiae Ser30 → Pro increased, Asp345 → Pro did not alter and Glu408 → Pro greatly decreased GA stability as measured by resistance to irreversible thermoinactivation relative to the wild-type enzyme. The Ser30 → Pro mutation was combined with other previously identified stabilizing mutations to examine whether combining such mutations ...


Suppressor Analysis Of The Protein Kinase Elm1p, An Enzyme Involved In The Regulation Of Yeast Morphology , Nicholas Patrick Edgington Jan 1997

Suppressor Analysis Of The Protein Kinase Elm1p, An Enzyme Involved In The Regulation Of Yeast Morphology , Nicholas Patrick Edgington

Retrospective Theses and Dissertations

Saccharomyces cerevisiae is an ascomycete that has several developmental options, with concomitant morphological changes, that depend on cell type and nutrient status. In response to these signals, vegetative Saccharomyces cerevisiae cells assume one of two distinct morphologies, the yeast form or the filamentous form. Molecules affecting this morphogenetic response were identified by analyzing transposon-induced or spontaneous suppressor mutations that restore yeast form growth to a strain that constitutively exhibits filamentous form characteristics owing to mutation of the protein kinase Elm1p. One transposon-tagged mutation was isolated and is allelic to the negative regulatory subunit Bcy1p, thus activation of cAMP dependent protein ...


Characterization Of Membrane-Associated Nuclease Activity In Mycoplasma Pulmonis , Karalee Jean Jarvill-Taylor Jan 1996

Characterization Of Membrane-Associated Nuclease Activity In Mycoplasma Pulmonis , Karalee Jean Jarvill-Taylor

Retrospective Theses and Dissertations

Mycoplasmas are deficient in numerous biosynthetic pathways. Consequently, growth is dependent upon the acquisition of structural and macromolecular precursors from the environment. These precursors include phospholipids, fatty acids and nucleotide bases. DNases and RNases are postulated to be involved in the acquisition of nucleotides. The goals of this study were to determine if nuclease activities are universally associated with mycoplasmas, characterize the nucleases by subcellular distribution and apparent molecular weight, and to identify and analyze the nuclease gene(s) in Mycoplasma pulmonis;Nuclease activity was present in all mycoplasma species tested. The level of activity did not correlate with virulence ...


Integration Specificity Of The Retrovirus-Like Transposable Element Ty5 Of Saccharomyces , Sige Zou Jan 1996

Integration Specificity Of The Retrovirus-Like Transposable Element Ty5 Of Saccharomyces , Sige Zou

Retrospective Theses and Dissertations

The retrotransposons and retroviruses display a strong preference in selecting integration sites. The molecular mechanisms underlying this process, however, are largely unknown. I have characterized a new retrotransposon, called Ty5, in diverse strains of Saccharomyces cerevisiae and related species. A functional element, Ty5-6p, was identified from S. paradoxus. To investigate Ty5 target preference, the locations of 13 native insertions were determined in S. cerevisiae and S. paradoxus. Twelve were found near the telomeres and the mating locus HMR. To determine if this distribution is a consequence of targeted integration, a Ty5 transposition assay was developed in S. cerevisiae using Ty5-6p ...


Sequence Variation Characteristics Of D-Loop, Rrna, Trna, And Polypeptide Coding Region Of Bovine Mitochondrial Dna , Jianming Wu Jan 1995

Sequence Variation Characteristics Of D-Loop, Rrna, Trna, And Polypeptide Coding Region Of Bovine Mitochondrial Dna , Jianming Wu

Retrospective Theses and Dissertations

By cloning and sequencing the mtDNA of cows from different maternal lineages, it was found that multiple heteroplasmic mtDNA states within the D-loop and rRNA genes of mtDNA existed extensively in Holstein cattle. Meanwhile, the nature and transmission of mtDNA were investigated. The mtDNA population in a specific animal was found to be a mixture of different genotypes. Because heteroplasmy was observed frequently and seemingly is persistent, selected amplification of specific mtDNA molecules may not be a part of mtDNA inheritance. At same time, four hypervariable sites were located at nt 169, 216, and 1594 between nt 352 to 364 ...


Telomeric Nucleic Acids: C-Strand Structure And A Telomerase Rna Mutant , Shawn Cameron Ahmed Jan 1994

Telomeric Nucleic Acids: C-Strand Structure And A Telomerase Rna Mutant , Shawn Cameron Ahmed

Retrospective Theses and Dissertations

Telomeres, the ends of linear chromosomes, are composed of simple tandem repeats which are usually G·C rich. Telomeres are essential for chromosome stability, organizing the nuclear architecture and ensuring complete replication of the chromosomal terminus. To understand how telomeres carry out these fundamental cellular roles, one must understand the structural and dynamic properties of telomeric repeat sequences. Structural and genetic approaches were taken to learn more about telomeric nucleic acids;The structural portion of my research concerned an unusual DNA structure formed by the C-rich strand of telomeric DNA. Telomeric C-strand sequences form non-Watson-Crick structures in supercoiled plasmids at ...


Genetic Analysis Of Pseudohyphal Growth In The Budding Yeast Saccharomyces Cerevisiae , Melissa Jo Blacketer Jan 1994

Genetic Analysis Of Pseudohyphal Growth In The Budding Yeast Saccharomyces Cerevisiae , Melissa Jo Blacketer

Retrospective Theses and Dissertations

The budding yeast Saccharomyces cerevisiae is a dimorphic organism that can assume either a yeast-like or pseudohyphal form. Nitrogen limitation induces pseudohyphal growth, which is characterized by branched chains of elongated cells. Pseudohyphal cells can grow invasively in agar medium, whereas yeast-like cells do not. To identify factors involved in morphologic differentiation, S. cerevisiae mutants exhibiting a constitutive cell elongation morphology were isolated. Genetic analysis identified 28 recessive and 2 semi-dominant mutations that cause abnormal morphology, and placed these in 14 distinct gene loci, termed ELM, for ELongated Morphology. Many elm mutations cause multiple aspects of pseudohyphal growth, and thus ...


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 ...


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 ...


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 ...