Genetics and Genomics Commons^™

Cohesion Without Cohesins In Drosophila Meiosis, Avik Mukherjee

Doctoral Dissertations

Meiosis is essential for sexual reproduction. The proper segregation of chromosomes in meiosis requires multiple functions of a multi-subunit protein complex known as cohesin. Cohesin forms a ring around duplicated sister chromatids and prevents them from separating prematurely. In Drosophila, mitotic cohesin is composed of four subunits: SMC1 and SMC3 and SCC1/RAD21 and SCC3/SA and are required for mitotic cohesion. However, RAD21 is dispensable for meiotic cohesion and although SMC1 and SMC3 are present on meiotic centromeres and on synapsed chromosome arms, as expected, their functions in meiosis remain poorly characterized. Moreover, unlike in other eukaryotes in which screens for …

Whole Genome Sequencing As A Tool For Identifying Phenotypic Properties And Underlying Genetic Mechanisms In Staphylococcus Pseudintermedius, Matthew C. Riley

Doctoral Dissertations

Staphylococcus pseudintermedius is a Gram-positive bacterial opportunistic pathogen commonly associated with dermal infections in canines, but capable of causing serious disease in other species. Reports of human infections caused by S. pseudintermedius along with an increase in resistance to multiple antibiotics highlights the importance of this organism. Whole genome sequencing can allow large scale investigation of genetic mechanisms underlying phenotypic properties that contribute to the expansion of successful S. pseudintermedius clonal lineages.

The increase in multidrug and methicillin-resistant S. pseudintermedius (MRSP) may result from horizontal transfer of genetic material between bacterial isolates, yet is thought to be rare in Staphylococci …

Tetrameric Photosystem I: From Initial Discovery And Characterization In Chroococcidiopsis Sp. Ts-821 To Exploration Of Its Distribution And Understanding Of Its Significance In Cyanobacteria, Meng Li

Doctoral Dissertations

Photosystem I (PSI) forms trimeric complexes in most characterized cyanobacteria. We had reported the tetrameric form of PSI in the unicellular cyanobacterium, Chroococcidiopsis sp. TS-821 (TS-821). Using Cryo-EM, a 3D model of the PSI tetramer structure at 11.5 [Angstrom] resolution was obtained and a 2D map within the membrane plane of at 6.1 [Angstrom]. In contrast to the three-fold symmetry in trimeric PSI crystal structure from T. elongatus, two different inter-monomer interactions involving PsaLs are found in the PSI tetramer. Phylogenetic analysis based on PsaL protein sequences shows that TS-821 is closely related to heterocyst-forming cyanobacteria. Additionally, this tetrameric …

Expansion Of And Reclassification Within The Family Lachnospiraceae, Kelly N. Haas

Doctoral Dissertations

Many of the taxa in the family Lachnospiraceae are currently misclassified as Clostridium spp. Here attempt to rectify many of these issues, beginning with an in-depth genomic and physiologic analysis of Clostridium methoxybenzovorans, culminating in the assertion that is a heterotype of Clostridium indolis, followed by reclassification of the broader group in which this organism resides. We propose two novel genera, Lacriformis and Enterocloster, to reclassify this clade, this includes reclassification of Clostridium sphenoides, Clostridium indolis, Clostridium saccharolyticum, Clostridium celerecrescens, Clostridium xylanolyticum, Clostridium algidixylanolyticum, Clostridium aerotolerans, Clostridium amygdalinum, and …

Improving Metabolic Engineering And Characterization Of Clostridium Thermocellum For Improved Cellulosic Ethanol Production, Beth Alexandra Papanek

Doctoral Dissertations

Biofules are an important option for humanity to move away from its dependence on fossil fuels. Transitioning from food crops to lignocellulosic alternatives for the production of biofuels is equally important. Most commonly, biofuels are produced using a crop such as corn or soybeans to feed sugars to the yeast, Saccharomyces cerevisiae for the fermentation of ethanol. Lignocellulosic biofuel production would eliminate the need for food crops and transition to biomass such as switchgrass, poplar, or corn stover. Currently, lignocellulosic biofuel production is limited primarily because of the cost of converting the biomass to fermentable sugars than can then be …

Pigment Dispersing Factor: Transcriptional Regulation And Its Role In Metabolism In Drosophila Melanogaster, Sudershana Nair

Doctoral Dissertations

Almost all living organisms have circadian clocks coordinating physiology and behavior, and an innate molecular clock drives rhythmic changes by integrating environmental and metabolic stimuli to generate 24 hour timing. Drosophila melanogaster has proved to be an excellent model organism with a well-characterized circadian clock and the neural circuits underlying clock have been intensely investigated. The neuropeptide pigment-dispersing factor (PDF) plays an essential role in maintaining circadian rhythmicity and synchronizes circadian clock neurons. However, the regulation of Pdf has been a black box with no known protein identified that directly regulates it, and its role in metabolism hasn’t been looked …

Novel Advancements For Improving Sprout Safety, Kyle S. Landry

Doctoral Dissertations

All varieties of bean sprouts (mung bean, alfalfa, broccoli, and radish) are classified as a “super-food” and are common staples for health conscious consumers. Along with the proposed health benefits, there is also an inherent risk of foodborne illness. When sprouts are cooked, there is little risk of illness. The purpose of this dissertation was to explore novel techniques to minimize or prevent the incidence of foodborne illness associated with the consumption of sprouts. Three areas were investigated: 1) the use of a biocontrol organism, 2) the use of a novel spontaneous carvacrol nanoemulsion, and 3) the influence of the …

Genome Engineering In Mammalian Cells By Flp And Cre Dna Recombinase Variants, Riddhi Shah

Doctoral Dissertations

Genome engineering relies on DNA modifying enzymes that are able to locate a DNA sequence of interest and initiate a desired genome rearrangement. Currently, the field predominantly utilizes site-specific DNA nucleases that depend on the host DNA repair machinery to complete a genome modification task. We show here that genome engineering approaches that employ self-sufficient, versatile site-specific DNA recombinase Flp and Cre can be developed into promising alternatives. We demonstrate that a Flp variant evolved to recombine an FRT-like sequence FL-IL10A, which is located upstream of the human interleukin-10 gene, can target this sequence in the model setting and native …

In Silico Driven Metabolic Engineering Towards Enhancing Biofuel And Biochemical Production, Richard Adam Thompson

Doctoral Dissertations

The development of a secure and sustainable energy economy is likely to require the production of fuels and commodity chemicals in a renewable manner. There has been renewed interest in biological commodity chemical production recently, in particular focusing on non-edible feedstocks. The fields of metabolic engineering and synthetic biology have arisen in the past 20 years to address the challenge of chemical production from biological feedstocks. Metabolic modeling is a powerful tool for studying the metabolism of an organism and predicting the effects of metabolic engineering strategies. Various techniques have been developed for modeling cellular metabolism, with the underlying principle …

Characterizing Early-Life Microbiome Functionality In Premature Infant Gut By A Metaproteomics Approach, Weili Xiong

Doctoral Dissertations

Microbes inhabit all parts of human body that are exposed to the environment and their interactions with human host mutually benefit each other and play significant roles in human health and diseases. The human gastrointestinal tract harbors the largest population of the microbiota and has gained broad research attention and efforts over the past decade. Colonization of the gut by microbes begins at birth and this early-life bacterial establishment can impact infants’ health and even the human health and lifestyle across an entire life span. Recent studies on community structure and composition of infant gut microbiota have revealed the species …

Computational Identification Of Terpene Synthase Genes And Their Evolutionary Analysis, Qidong Jia

Doctoral Dissertations

Terpenoids, the largest and most structurally and functionally diverse class of natural compounds on earth, are mostly synthesized by plants to be involved in various plant environment interactions. Some terpenoids are classified as primary metabolites essential for plant growth and development. Terpene synthases (TPSs), the key enzymes for terpenoid biosynthesis, are the major determinant of the tremendous diversity of terpenoid carbon skeletons. The TPS genes represent a mid-size family of about 30-100 functional genes in almost all major sequenced plant genomes. TPSs are also found in fungi and bacteria, but microbial TPS genes share low levels of sequence similarity and …

Biochemistry And Evolution Of The Phytohormone-Methylating Sabath Methyltransferase In Plants, Minta Chaiprasongsuk

Doctoral Dissertations

Known members of Phytohormone-methylating compounds are plant synthesis compounds that serve as attractants of other living organisms beneficial to the plants or as defense against other biotic as well as abiotic agents. To increase their fitness and survival in a stressful environment plants produce distinct sets of phytohormone-methylating compounds. Plant genomes can encode the necessary enzymes to acquire the ability to make new specialized compounds during evolution. This dissertation aims to investigate the biochemical and biological functions and evolution of SABATH genes in different lineages of plants. Black cottonwood, Brachypodium and Norway spruce genome were used as the model for …

Evolvability Of The Skull: A Study Of Genetic Basis And Integration In The Teleost Craniofacial Skeleton, Yinan Hu

Doctoral Dissertations

As the field of evolutionary biology pivots away from a gene-centric view of how adaptive evolution proceeds, renewed emphasis is placed on the origin of phenotypic variation. Understanding the developmental processes that underlie the production of novel traits, and how they might influence evolvability, is considered a primary goal in the on-going “extended evolutionary synthesis”. The following dissertation explores these questions in the context of adaptive radiations in fish, with a focus on morphological variation in the craniofacial skeleton. Specifically, the first chapter investigates the genetic and developmental basis of shape (co-)variation in the feeding apparatus of African cichlid fishes, …

Bacteriophage: Bioengineered Bacterial Detection And Applications, Samuel D. Alcaine

Doctoral Dissertations

Bacteria are ubiquitous and vital constituents of our environment, our foods, and our bodies. A small percentage of this vast, microbial population is pathogenic to humans, but represents a significant burden on public health. There is a current public health focus on two subgroups: foodborne pathogenic bacteria and antibiotic resistance bacteria. A key challenge for public health is the rapid identification of these bacteria to prevent their consumption and to ensure proper treatment for infections. This challenge calls for the development of novel, low-cost diagnostics that combine sensitivity and accuracy with speed and ease-of-use. Bacteriophages represent rapid, readily targeted, and …

Size Specific Transfection To Mammalian Cells By Micropillar Array Electroporation, Yingbo Zu

Doctoral Dissertations

Electroporation serves as a promising non-viral gene delivery approach, while its current configurations carry drawbacks associated with high-voltage electrical pulses and heterogeneous treatment on individual cells. Here, we developed a new micropillar array electroporation (MAE) platform to advance the delivery of plasmid DNA and RNA to mammalian cells. By introducing well-patterned micropillar array on the electrode surface, the number of pillars each cell faces varies with its cell membrane surface area, despite their large population and random locations. In this way, cell size specific electroporation is conveniently done and contributed to a 2.5~3 fold increase on plasmid DNA transfection and …