Bacteria Commons^™

Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

We report development of a genetic system for making targeted gene knockouts in Clostridium thermocellum, a thermophilic anaerobic bacterium that rapidly solubilizes cellulose. A toxic uracil analog, 5-fluoroorotic acid (5-FOA), was used to select for deletion of the pyrF gene. The ΔpyrF strain is a uracil auxotroph that could be restored to a prototroph via ectopic expression of pyrF from a plasmid, providing a positive genetic selection. Furthermore, 5-FOA was used to select against plasmid-expressed pyrF, creating a negative selection for plasmid loss. This technology was used to delete a gene involved in organic acid production, namely pta, which encodes …

The Effects Of Host Physiological Conditions On The Expression Of Icsp In Shigella Flexneri, Karen Levy, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a gram-negative bacterium capable of causing diarrhea and dysentery known as shigellosis. It is estimated there are 167.4 million shigellosis episodes throughout the world each year causing 1.1 million deaths. Shigella invades cells in the lower intestine through an induced phagocytosis. Once in the cytoplasm, bacteria move from one cell to another using actin-based motility. The Shigella outer membrane protease IcsP regulates actin-based motility and cell-to-cell spread by cleaving the actin assembly protein IcsA from the bacterial cell surface. We hypothesize that IcsP may serve additional functions during infection. By examining which environmental signals trigger icsP expression, …

Characterization Of The Omptin Protease, Ompt, In Escherichia Coli, Amanda Yates, Eun-Hae Kim, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Omptins are outer membrane proteases found in gram negative bacteria that cause diseases in humans, such as pathogenic Escherichia coli, Shigella flexneri, Salmonella typhimurium, and Yersinia pestis. Bacterial species that express omptins cause diseases such as highly fatal plague and severe diarrhea and dysentery. The genes that encode these proteases are ompT, icsP, pgtE, and pla, respectively. These proteases are highly related in structure and share approximately 50% sequence identity. In S. flexneri, IcsP has been shown to cleave a key virulence determinant, IcsA (Egile et al., 1997). IcsA recruits host actin and allows for intracellular movement within host cells …

Antimicrobial Activity Screening Of Recombinant And Synthetic Varasin A Defensin From The Hard Tick Dermacentor Variabilis Against Various Bacteria, Julia A. Sharp

Biological Sciences Theses & Dissertations

Hematophagous arthropods, such as ticks and mosquitoes, rely on their innate immune system for defense against pathogens ingested in a blood meal as well as those acquired through injury. In response to pathogen recognition, the production of antimicrobial peptides, such as defensin, is typically upregulated. Varisin, a defensin, is thought to be a key component in the immunocompetence of the hard tick D. variabilis against Borrelia burgdorferi, the causative agent of Lyme disease. To study the antimicrobial effects of varisin, recombinant varisin was expressed by both insect cells and E coli. Purification of the protein followed by enterokinase treatment yielded …

Downstream Pathway Activation In Cultured Hepatocytes Following Treatment With Francisella Tularensis, Nicole Wakefield

Undergraduate Theses and Capstone Projects

Francisella tularensis, the intracellular pathogen that causes tularemia, was investigated to determine how it infects and replicates within mammalian hepatocytes. To date, it has been shown that hepatocytes can be infected by F. tularensis and that this causes considerable change in the protein phosphorylation state of several vital signaling molecules within the host cell. Protein kinase pathways can be mapped as signatures of infection. Hepatocytes tend to be susceptible to infection by F. tularensis, thus stimulating internal signaling. The Francisella strains used were selected with the goal of producing a model that can be used to elucidate the cell signaling …

Molecular Screening Of Azoreductase Gene And Its Activity In Human Intestinal Bacteria, Syndia S. Todd

McCabe Thesis Collection

The intestinal microflora are capable of performing a wide variety of metabolic transformations. "The intestinal bacteria can enhance the function of the entire gastrointestinal tract, protect against pathogenic, maintain the vital chemical balance of the gastrointestinal system, and produce needed vitamins and hormones" (http;//www.upwardquest.com/crit2.html). Some of the products of this metabolism have been associated with carcinogenic processes, such as cancer, tumor formation, gastrointestinal disease, and infections. The ability of human intestinal microbes to interact with metabolites directly or after recirculation may contribute toward different toxicological disorders and disease.

The purpose of this study is to characterize and isolate the azoreductase …

Metal-Resistance Genetically Engineered Bacteria, Sylvia Daunert, Donna Scott, Sridhar Ramanathan

KWRRI Research Reports

Bacterial-based electrochemical and optical sensing systems that respond in a highly selective and sensitive manner to antimonite and arsenite have been developed. This was accomplished by using genetically engineered bacteria bearing one of two plasmids constructed for our studies. The first plasmid, pBGD23, contains the operator/promoter region (O/P) and the gene of the ArsR protein from the ars operon upstream from the β-galactosidase gene. In the absence of antimonite/arsenite, ArsR binds to the 0/P site and prevents the transcription of the genes for ArsR and β-galactosidase, thus blocking expression of these proteins. When antimonite or arsenite is present in the …

Movement Of Bacteria Through Macropores To Ground Water, M. Scott Smith, Grant W. Thomas, Robert E. White

KWRRI Research Reports

Effects of soil type, flow rate, antecedent soil moisture and other factors on transport of E. coli through soils was measured on disturbed and intact columns 20 cm in diameter by 25 to 30 cm in depth. Added E. coli were distinguished from indigenous microbes using an antibiotic resistance marker. Transport of Cl^- and ³H₂O was also measured. Up to 96 percent of the bacteria irrigated onto the surface of intact columns were recovered in the effluent. Soil structure appeared to be related to the extent of transport. Columns prepared from mixed, repacked soil were much …

The Overwintering Of The Fireblight Bacteria Outside Of Living Tissue In Utah, Sherman V. Thomson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The purpose of this investigation was to determine the sites of overwintering of Erwinia amylovora (Burrill) Winslow et al. outside of living tissue in Utah. Bacterial isolates were taken from the surfaces of blighted twigs and mummified fruits remaining on the tree; also from internal dead tissue of blighted twigs and mummies, and from soil beneath blighted trees. Each isolate was tested for agglutination with antiserum specific for Erwina amylovora, susceptibility to Erwinia amylovora bacteriophage, production of symptoms in pear seedlings and development of bacterial ooze on green pears.

Both virulent and avirulent isolates were taken from all sites tested. …