Molecular Genetics Commons^™

Snd1-Targeted Gene Therapy For Hepatocellular Carcinoma, Bryan D. Mckiver

Theses and Dissertations

Staphylococcal nuclease and tudor-domain containing 1 (SND1) is an oncogene for a wide variety of cancers, including hepatocellular carcinoma (HCC). SND1 is a multifunctional protein regulating gene expression of proto-oncogenes and tumor suppressor genes, making SND1 a prime target for developing cancer therapeutics. This notion is especially attributed to HCC as most patients are diagnosed in advanced stages and the therapeutic options available for these patients are severely limited. In this study, we evaluated the therapeutic potential of a replication-defective adenovirus vector delivering SND1 shRNA (Ad.SND1sh) to human HCC cell lines, HepG3, HuH-7, and Hep3B. Adenovirus infection in HCC cells …

Processing Of 3′-Blocked Dna Double-Strand Breaks By Tyrosyl-Dna Phosphodiesterase 1, Artemis And Polynucleotide Kinase/ Phosphatase, Ajinkya S. Kawale

Theses and Dissertations

DNA double-strand breaks (DSBs) containing unligatable termini are potent cytotoxic lesions leading to growth arrest or cell death. The Artemis nuclease and tyrosyl-DNA phosphodiesterase (TDP1) are each capable of resolving protruding 3′-phosphoglycolate (PG) termini of DNA double-strand breaks (DSBs). Consequently, a knockout of Artemis and a knockout/knockdown of TDP1 rendered cells sensitive to the radiomimetic agent neocarzinostatin (NCS), which induces 3′-PG-terminated DSBs. Unexpectedly, however, a knockdown or knockout of TDP1 in Artemis-null cells did not confer any greater sensitivity than either deficiency alone, indicating a strict epistasis between TDP1 and Artemis. Moreover, a deficiency in Artemis, but not TDP1, resulted …

Chemical Topping Burley Tobacco, Mitchell D. Richmond

Theses and Dissertations--Plant and Soil Sciences

The act of topping tobacco (Nicotiana tabacum L.) involves the removal of the terminal bud or inflorescence of the tobacco plant. This practice ordinarily is accomplished by manually removing the top of each tobacco plant in an entire field which is labor intensive and costly. Chemical topping utilizes sucker control products to inhibit the terminal bud and axillary bud growth without manually removing the top of the tobacco plant. There were several research objectives in order to determine the utility of a chemical topping system: 1) determine if burley tobacco could be chemically topped with currently registered suckercide products …

Floral Symmetry Genes Elucidate The Development And Evolution Of Oil-Bee Pollinated Flowers Of Malpighiaceae And Krameriaceae, Farahnoz N. Khojayori

Theses and Dissertations

Specialization on insect and animal pollinators is thought to be the driving force for the evolution of floral traits. Specifically in the New World (NW), the oil-bee pollination syndrome has led to the convergence of floral characters in two distantly related families of core eudicots, Malpighiaceae and Krameriaceae. Both families display a flag-like structure that establishes a zygomorphic flower and floral oil rewards in epithelial elaiophores. These traits work concomitantly to attract and reward female oil-bees that help fertilize these flowers and in return receive oils. The underlying genetics of floral zygomorphy were studied in several clades of core eudicots, …

Tetratricopeptide 39c (Ttc39c) Is Upregulated During Skeletal Muscle Atrophy And Is Necessary For Muscle Cell Differentiation, Caleb Hayes

UNF Graduate Theses and Dissertations

Ttc39c has been identified as a novel gene in skeletal muscle that is upregulated in response to neurogenic atrophy in mice. Quantitative PCR and Western blot analysis confirmed that Ttc39c is expressed in both proliferating and differentiated muscle cells. Furthermore, comparison of Ttc39c expression in undifferentiated and differentiated C₂C₁₂ cells demonstrated that Ttc39c levels peak in early differentiation, but decreases as cells become fully differentiated myotubes. The transcriptional regulation of Ttc39c was examined by cloning promoter fragments of the gene and fusing it with the SEAP reporter gene. The Ttc39c reporter gene constructs were transfected into muscle …

Estimating The Heritability Of Thermal Tolerance In Acropora Cervicornis And The Physiological Basis Of Adaptation That Correlates To Survival At Elevated Temperatures, Kelsey L. Yetsko

UNF Graduate Theses and Dissertations

Human activities have substantially increased the atmospheric concentrations of greenhouse gases, resulting in warmer ocean temperatures that are having a negative impact on reef corals, which are highly susceptible to changes in temperature. Understanding the degree to which species vary in their tolerance to elevated temperatures and whether this variation is heritable is important in determining their ability to adapt to climate change. In order to address this, Acropora cervicornis fragments from 20 genetically distinct colonies were kept at either ambient or elevated temperatures, and mortality was monitored for 26 days. Heritability of thermal tolerance was estimated using a clonal …

การจำแนกลักษณะทางโมเลกุลของเชื้อ Escherichia Coli ที่สร้างเอนไซม์ Extended-Spectrum Beta-Lactamases ที่แยกได้จากสุกรและประชากรในอำเภอเมือง จังหวัดลำพูน, จักรพงษ์ สีนามะ

Chulalongkorn University Theses and Dissertations (Chula ETD)

ปัจจุบันพบเชื้อ Escherichia coli ที่สร้างเอนไซม์ extended-spectrum beta-lactamase (ESBL) แพร่กระจายในชุมชนอย่างกว้างขวาง เชื้อแบคทีเรียดังกล่าวสามารถพบได้ว่าเป็นเชื้อประจำถิ่นในลำไส้ของคน และสัตว์ รวมไปถึงสามารถพบได้ในสิ่งแวดล้อมทั่วไป ซึ่งอาจกลายเป็นแหล่งเก็บกักเชื้อแบคทีเรียดื้อสารต้านจุลชีพ ที่สามารถแพร่กระจายในชุมชนต่างๆ ทั่วโลก การศึกษาครั้งนี้ทำการจำแนกลักษณะทางโมเลกุลของเชื้อ E. coli ที่สร้างเอนไซม์ ESBL ซึ่งแยกได้จากสุกรในฟาร์มปศุสัตว์และประชากรในชุมชน ในพื้นที่อำเภอเมือง จังหวัดลำพูน เพื่อศึกษารูปแบบของยีนที่ควบคุมการสร้างเอนไซม์ ESBL และศึกษาหาความสัมพันธ์ของเชื้อในกลุ่มตัวอย่างทั้งสอง เชื้อ E. coli ที่สร้างเอนไซม์ ESBL จำนวนทั้งสิ้น 212 สายพันธุ์ แยกได้จากอุจจาระของสุกรและประชากรที่มีสุขภาพดี และทำการจำแนกยีนที่ควบคุมการสร้างเอนไซม์ ESBL ได้แก่ ยีน blaCTX-M ยีน blaTEM และยีน blaSHV ด้วยเทคนิค multiplex-PCR และเทคนิค whole genome sequencing และศึกษาความสัมพันธ์ของเชื้อ E. coli ที่สร้างเอนไซม์ ESBL ที่แยกได้จากทั้งสองกลุ่มตัวอย่างด้วยเทคนิค multilocus sequence typing ผลการศึกษาพบว่าเชื้อ E. coli ที่สร้างเอนไซม์ ESBL ส่วนใหญ่เป็นเชื้อดื้อยาหลายขนาน และมักดื้อยากลุ่ม fluoroquinolones ยากลุ่ม aminoglycosides และยา trimethoprim/sulfamethoxazole ในขณะที่เชื้อทุกสายพันธุ์ดื้อต่อยา ampicillin ยา ceftriaxone และยา cefotaxime ยีนที่ควบคุมการสร้างเอนไซม์ ESBL ที่พบมากที่สุดได้แก่ ยีน blaCTX-M ซึ่งพบได้ 95.75% รองลงมาคือยีน blaTEM และยีน blaSHV พบได้ 62.73% และ 2.40% ตามลำดับ โดย subgroup ของยีนที่ควบคุมการสร้างเอนไซม์ ESBL ที่พบมากที่สุด ได้แก่ ยีน blaCTX-M-55 …

Comparison Between Listeria Sensu Stricto And Listeria Sensu Lato Strains Identifies Novel Determinants Involved In Infection, Jakob Schardt, Grant Jones, Stefanie Müller-Herbst, Kristina Schauer, Sarah E. F. D'Orazio, Thilo M. Fuchs

Microbiology, Immunology, and Molecular Genetics Faculty Publications

The human pathogen L. monocytogenes and the animal pathogen L. ivanovii, together with four other species isolated from symptom-free animals, form the "Listeria sensu stricto" clade. The members of the second clade, "Listeria sensu lato", are believed to be solely environmental bacteria without the ability to colonize mammalian hosts. To identify novel determinants that contribute to infection by L. monocytogenes, the causative agent of the foodborne disease listeriosis, we performed a genome comparison of the two clades and found 151 candidate genes that are conserved in the Listeria sensu stricto species. Two factors were …

Unseen Science: Modern Discoveries Too Far Away Or Tiny For Human Eyes, Lucy Huang

Capstones

As science has progressed, scientists have realized that evidence goes beyond the realms of physical sight. Whether it is too small or difficult to find, scientists have developed different ways to get around this problem. We see this in cancer genomics and in extrasolar planetary research. Scientists use what they know and what they measure to validate their work.

https://lucy-huang-9tge.squarespace.com/

Mechanistic Insights Into The Regulation Of Mitochondrial Fission By Cyclin C, Vidyaramanan Ganesan, Katrina F Cooper, Randy Strich

Rowan-Virtua School of Osteopathic Medicine Departmental Research

Cyclin C is a component of the mediator complex of RNA polymerase II that localizes to the nucleus under normal conditions. In response to stress, cyclin C translocates to the cytosol and mitochondria and mediates stress‐induced mitochondrial fission and apoptosis. The molecular mechanisms by which cyclin C induces mitochondrial fission are unknown. Using in vitro experimental approaches, we sought to investigate the mechanistic basis of cyclin C mediated mitochondrial fission.

Effects Of Crude Oil On Tumor Suppressor P53 Polymorphisms In Laboratory-Exposed Atlantic Killifish, Fundulus Heteroclitus, Allison Margaret Nadler

Seton Hall University Dissertations and Theses (ETDs)

Polycyclic aromatic hydrocarbons (PAHs), constituents of crude oil, are implicated as a potent source of adverse toxicological effects on living organisms. To model the effects of PAHs in response to environmental oil spill disasters a species of killifish (Fundulus heteroclitus) was captured and exposed to crude oil in a laboratory-controlled setting. Over a period of 7 days, fish were dosed with crude oil by gavage, culled, and organs were harvested for analysis. Excitation-emission matrix spectroscopy (EEMS) of gall bladder tissue homogenates was used to verify exposure. Effects of PAHs on the p53 gene were evaluated as an indicator …

A Neuroprotective Role For Mir-1017, A Non-Canonical Mirna, Matthew De Cruz

Master's Theses

miRNAs are post-transcriptional regulators of gene expression, with numerous being involved in neurobiology. Within the human genome a quarter of the identified miRNA loci derive from a class of miRNAs termed tailed mirtrons. Despite the identification of this large population of miRNA, no functional studies have been conducted to identify their role. In this study we examined the highly expressed and deeply conserved Drosophila 3’ tail mirtron, miR-1017, as a candidate to elucidate tailed mirtron functionality. We identified acetylcholine receptor transcripts, Da5 and Da2, as bona fide targets for miR-1017. Interestingly, Da2 is also the host transcript for miR-1017. We …

Ablation Of An Ovarian Tumor Family Deubiquitinase Exposes The Underlying Regulation Governing The Plasticity Of Cell Cycle Progression In Toxoplasma Gondii, Animesh Dhara, Rodrigo De Paula Baptista, Jessica C. Kissinger, Ernest Charles Snow, Anthony P. Sinai

Microbiology, Immunology, and Molecular Genetics Faculty Publications

The Toxoplasma genome encodes the capacity for distinct architectures underlying cell cycle progression in a life cycle stage-dependent manner. Replication in intermediate hosts occurs by endodyogeny, whereas a hybrid of schizogony and endopolygeny occurs in the gut of the definitive feline host. Here, we characterize the consequence of the loss of a cell cycle-regulated ovarian tumor (OTU family) deubiquitinase, OTUD3A of Toxoplasma gondii (TgOTUD3A; TGGT1_258780), in T. gondii tachyzoites. Rather than the mutation being detrimental, mutant parasites exhibited a fitness advantage, outcompeting the wild type. This phenotype was due to roughly one-third of TgOTUD3A-knockout (TgOTUD3A-KO) tachyzoites exhibiting deviations from endodyogeny …

Mechanism Of Transcription Anti-Termination In Human Mitochondria., Hauke S Hillen, Andrey V Parshin, Karen Agaronyan, Yaroslav I Morozov, James J Graber, Aleksandar Chernev, Kathrin Schwinghammer, Henning Urlaub, Michael Anikin, Patrick Cramer, Dmitry Temiakov

Rowan-Virtua School of Osteopathic Medicine Departmental Research

In human mitochondria, transcription termination events at a G-quadruplex region near the replication origin are thought to drive replication of mtDNA by generation of an RNA primer. This process is suppressed by a key regulator of mtDNA-the transcription factor TEFM. We determined the structure of an anti-termination complex in which TEFM is bound to transcribing mtRNAP. The structure reveals interactions of the dimeric pseudonuclease core of TEFM with mobile structural elements in mtRNAP and the nucleic acid components of the elongation complex (EC). Binding of TEFM to the DNA forms a downstream "sliding clamp," providing high processivity to the EC. …

An Undergraduate Laboratory Manual For Analyzing A Crispr Mutant With A Predicted Role In Regeneration, Susan Walsh, Ashley Becker, Paxton S. Sickler, Damian G. Clarke, Erin Jimenez

Faculty Publications

Exposing students to undergraduate research has reportedly improved students’ development of knowledge and skills in the laboratory, self-efficacy, satisfaction with their research, retention, and perseverance when faced with obstacles. Furthermore, utilizing authentic course-based undergraduate research experiences (CUREs) includes all students enrolled in the class, giving those who may not otherwise have access to an independent undergraduate research project an opportunity to engage in the scientific process in context of an original, unanswered question. In the fall of 2016, second semester introductory biology students conducted a semester-long research project on the transcription factor Lin28a to determine the effect of Lin28a on …

The Impact Of Co2-Related Ocean Acidification On The Molecular Regulation Of Shell Development In The Eastern Oyster (Crassostrea Virginica)., Mackenzie L. Richards

LSU Master's Theses

Eastern oysters (Crassostrea virginica), native to the Gulf of Mexico, are keystone species in estuarine ecosystems and are economically valued. Current research indicates that ocean acidification adversely affects the physiology and morphology of larval oysters, but the molecular mechanisms of this impact remain unstudied. Ocean acidification is contributed to by elevated atmospheric CO₂ due to increased anthropogenic activities, causing heightened partial pressure of CO₂ (pCO₂), and eutrophication from land-based runoff in the Gulf. The objective of this work was to determine the genomic response of the eastern oyster in Louisiana to simulated ocean acidification. …

The Role Of Histone H2a.Z Abundance In Modulating Responses To Phosphorus Deficiency In Rice, Sara Zahraeifard

LSU Doctoral Dissertations

Histone variants contribute to chromatin complexity by creating specialized nucleosomes. Here, to investigate the pattern of H2A.Z histone variant distribution and its role in modulating gene expression in rice (Oryza sativa L.), we performed genome-wide profiling of ARP6-dependent H2A.Z deposition in rice seedlings. We demonstrated that under controlled conditions, the majority of H2A.Z deposition is within protein-coding genes (PCG), and the most highly expressed genes are enriched with H2A.Z at their 5’ ends near the transcription start site (TSS), whereas the medium- to low-expressed genes contain H2A.Z across the entire gene. Based on H2A.Z deposition, we categorized genes into …

Virulence Phenotypes Of Rotylenchulus Reniformis: Evaluation Of Host Status Of Cotton And Utility Of Single Nucleotide Polymorphisms (Snps) For Identification, Churamani Khanal

LSU Doctoral Dissertations

Comparative reproduction and pathogenicity of reniform nematode (Rotylenchulus reniformis) populations derived from single-egg masses and collected form West Carroll (WC), Rapides (RAP), Morehouse (MOR), and Tensas (TEN) parishes in Louisiana were evaluated in microplot and greenhouse trials. Data from microplot trials showed significant differences among isolates of reniform nematode in both reproduction and pathogenicity on upland cotton (Gossypium hirsutum) cultivars Phytogen 499 WRF, Deltapine 1133 B2RF, and Phytogen 333 WRF. Across all cotton cultivars, MOR and RAP isolates had the greatest and the least reproduction values of 331.8 and 230.2, respectively. Reduction in plant dry weight, …

Using Competition Assays To Quantitatively Model Cooperative Binding By Transcription Factors And Other Ligands., Jacob Peacock, James B Jaynes

Department of Biochemistry and Molecular Biology Faculty Papers

BACKGROUND: The affinities of DNA binding proteins for target sites can be used to model the regulation of gene expression. These proteins can bind to DNA cooperatively, strongly impacting their affinity and specificity. However, current methods for measuring cooperativity do not provide the means to accurately predict binding behavior over a wide range of concentrations.

METHODS: We use standard computational and mathematical methods, and develop novel methods as described in Results.

RESULTS: We explore some complexities of cooperative binding, and develop an improved method for relating in vitro measurements to in vivo function, based on ternary complex formation. We derive …

Characterization Of A Novel Mitochondrial Plasmid In Brassica, Mackenzie Strehle

UCARE: Research Products

Possessing some of the largest and most complex genomes of any eukaryotic organelles, plant mitochondria are notorious for their rapidly rearranging genetic framework. In addition to containing a large and complex mitochondrial genome, the mitochondria of several plants in the genus Brassica have also been shown to contain an independent, self-replicating linear plasmid. Interestingly, the plasmid appears to be able to move independently between the cytoplasm and the mitochondria, and it can be paternally inherited, unlike the rest of the mitochondrial genome. The plasmid also has features similar to those of adenoviruses, including terminal inverted repeats and covalently bound proteins …