Biotechnology Commons^™

Dna Purification Process Optimization At Life Technologies Corporation, Trevor J. Shepherd

Master's Theses

This project focused on optimizing the plasmid DNA purification process at Life Technologies. These plasmids are designed to code for specialized proteins used by research universities, national laboratories, or research companies. Once cultivated and harvested, the plasmids must be analyzed for quality and quantity. The project is divided into improving three aspects of the process: 1) plasmid identification, 2) plasmid purity evaluation, and 3) process yield. Plasmid identification is now simpler, more robust and has zero ambiguity. Plasmid purity evaluation is now measured with computer software, which reduces user error and eliminates subjectivity. Using the nascent metrics provided by the …

Disocvering Ionic Liquid Resistant Genes, Bree Person, Douglass Higgins, Michael Thelen

STAR Program Research Presentations

: Plant biomass is a rich source of sugars that can be converted to biofuels by engineered microbes. However, because the lignocellulose in biomass is insoluble in aqueous conditions and recalcitrant to enzymatic degradation, thermochemical treatment is required to break apart the lignin and cellulose polymers before sugars can be released. The most effective chemicals for doing this are known as ionic liquids, which are salts that are molten at temperatures below 100° C. Although these solvents have many unique properties that are ideal for solubilizing lignocellulose, they have been found to inhibit the growth of bacterial strains used to …

Optimizing Electrode Design For Microbial Fuel Cells Used For Wastewater Treatment, Lindsay Nichols, John A. Hogan

STAR Program Research Presentations

Microbial fuel cells (MFC) utilize bacteria to generate an electrical current that can be used in the decomposition of sludge and human urine. In a MFC there is an anode (for oxidation of organic compounds), cathode (reduction of oxygen or carbon dioxide), and a proton exchange membrane (PEM, allows protons to migrate); reduction-oxidation reactions between the anode and cathode produce a measurable current. Bacteria that are found in sludge can be used to produce electrons in a voltaic cell, but optimizing conditions for harnessing the energy is crucial to making a MFC efficient. Research has shown that the ratios of …

Improving The Efficiency Of Homologous Gene Replacement By Disrupting Non-Homologous End Joining Pathway Gene Kusa In The Oleaginous Fungus Mortierella Alpina, Kathleen T. Krueger, Ziyu Dai Dr, Ugur Uzuner, Scott E. Baker Dr

STAR Program Research Presentations

Mortierella alpina , a oleaginous filamentous fungus, is one of industrial fungal strains known for the production of arachidonic acid. It is also of particular interest for hydrocarbon biofuel production since it is able to produce up to 50% of its mass in rich, long-chain polyunsaturated fatty acids [PUFA’s]. In addition to high fatty acid production, M. alpina like many other oleaginous fungi, already have mechanisms for accumulating significant concentrations of hydrophobic compounds making it a naturally equipped candidate to handle potential toxic concentrations of hydrocarbons. To date, three different transformation methods have been established for M. alpina that are …

Quantification And In Vitro Analysis Of Nanolipoproteins (Nlps) Containing Adjuvants, Purna Venkataraman, Craig Blanchette, Nicholas O. Fischer

STAR Program Research Presentations

Nanolipoprotein particles (NLPs) self-assemble into nanoscale structures that can be used as vaccines or drug delivery agents. Due to the nature of the NLPs, a variety of immune stimulating compounds or adjuvants can be readily incorporated into NLPs: a characteristic difficult to engineer into most other nanoscale platforms. In light of this, a method for quantifying the amount adjuvant actually incorporated into NLPs is a question of high importance. Through the use of reverse phase High-Performance Liquid Chromatography (HPLC) and an Evaporative Light Scattering Detector (ELSD), standard curves can be constructed by analyzing mixtures of NLP components of known concentration, …

Analyzing Environmental Microbes For Genomic Regions Promoting Ionic Liquid Tolerance In E. Coli, Ann Nguyen, Alison Richins, Thomas Rüegg, Steven Singer, Michael Thelen

STAR Program Research Presentations

Ionic liquids (ILs) are promising as solvents to increase the efficiency of biofuel production; however, ILs are toxic to microbes used in the fermentation of liquid fuels. To engineer IL resistant biofuel hosts, environmental bacteria were screened for tolerance, and these were used to create gene libraries to test in E. coli. Future characterization of these libraries using molecular techniques will be used to identify genes that contribute IL-tolerance to transformed microbes.

Efficacy Comparison Of Various Pcr Master Mixes And Thermal Cycling Parameters For Detection Of Brucella Abortus In Multiplex Assays, Kevin J. Testo, Jason A. Olivas, Alda C. Carrillo, Pejman Naraghi-Arani

STAR Program Research Presentations

Diagnostic testing for bioterrorism agents in food is essential for the safety and welfare of the general populace. Multiplex assays provide the means to test for multiple pathogen signatures with a single polymerase chain reaction (PCR) test. PCR, Luminex xMAP ® infrared fluorophore bead technology, and flow cytometry, multiplex diagnostic assays are incredibly specific and sensitive to highly conserved regions at various loci in a microbe’s genome. In order to increase the effectiveness of the PCR and the resulting fluorescent response of the beads, different PCR master mix commercial kits were compared to the standard AgPath RT-PCR mix. This study …

Enhanced Hydrocarbon Biodegradation Using Bioaugmentation With Biowish™-Aqua Fog, Michael Robert Lehrer

Master's Theses

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSH^TM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSH^TM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The …

Investigation Of Bacillus Subtilis As A Biopesticide Against Botrytis Cinerea, Kenneth K. Ng

Master's Theses

The objective of this thesis was to investigate BiOWiSH^TM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSH^TM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSH^TM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed …

Forward Osmosis Bag: Ground Testing The Prospect Of Using Alternative Water Sources For Drinking Water, Michaela Marie Brant, Ariella Yendler, Mona Hammoudeh, Michael T. Flynn

STAR Program Research Presentations

On the STS-135 Space Shuttle mission, to be launched July 8, 2011, a forward osmosis bag (FOB) study will be conducted. At NASA Ames this summer, the ground truth testing is being conducted for results comparisons. The FOB technology is derived from a commercial product, the X-Pack water filter. Forward osmosis operates by utilizing an established concentration gradient across a semi-permeable membrane to move water molecules from one side of the membrane to the other. This concept is exploited to harvest drinking water from grey water sources such as urine, sea water, or vehicle water. In this experiment, potassium chloride …

The Proteomic Response Of Ciona Intestinalis To Ocean Acidification And Acute Heat Stress: Impacts Of Global Climate Change In Coastal Marine Systems, Michael Dennis Dwyer Jr.

Biological Sciences

No abstract provided.

Pcr Primers For The Detection Of Propane And Butane-Oxidizing Microorganisms, Brian Jeremy Chan

Master's Theses

In an increasingly energy-hungry world, our capacity to meet the heightened energy demands of the future has become a pressing matter. The most urgent of these concerns are tied to the accessibility of petroleum. Various experts have proselytized both the imminent arrival of peak oil production rates and the ensuing decline of those rates thereafter. And to that end, the development of novel and advanced oil exploration methodologies has become almost as important as finding the sources of oil themselves.

The soils above petroleum reservoirs play host to various communities of alkane- oxidizing bacteria that can utilize the natural gas …

Growth Hormone Alters Components Related To Differentiation, Metabolism And Milk Synthesis And Secretion In Mac-T Cells, Tasha Lynn Johnson

Master's Theses

The mammary alveolar cell-T (MAC-T) cell line is able to uniformly differentiate and secrete casein proteins in response to dexamethasone, insulin and prolactin and is extensively used to study bovine mammary epithelial cell function. Growth hormone (GH) has been shown to increase milk protein synthesis both in vivo and in mammary cell models, and induce cytoskeletal rearrangement in 3T3 fibroblast cell line and a Chinese hamster ovary (CHO) cell line. Few studies have focused on identifying the mechanisms involved in differentiated MAC-T cells’ response to GH. We tested the hypothesis that MAC-T cells would respond directly to GH and that …

Consumers And Benefits Of Genetically Modified Vegetables, Megan Carter Judge

Master's Theses

With the adoption of biotechnology in many agricultural products with first-generation biotechnology traits such as increased pest resistance, greater herbicide resistance, and increased yields the growers have accepted them. The next wave of biotech crops have second-generation traits, such as improved nutrient content, extended shelf life, reduced pesticide and herbicide application (a consumer demanded trait), and better taste. Will these consumer benefits offset any concern that the consumer has regarding biotechnology? What are those benefits and how should the information be communicated to the consumer?

Three focus groups give insight to the proposed questions. The focus groups were done in …

Exposure To Ultraviolet Radiation Causes Proteomic Changes In Embryos Of The Purple Sea Urchin, Strongylocentrotus Purpuratus, Joseph Paul Campanale

Master's Theses

The amount of solar ultraviolet radiation (UVR, 290-400 nm) reaching Earth’s surface is increasing due to ozone depletion and global climate change. Embryos of the purple sea urchin, Strongylocentrotus purpuratus, provide an ideal system for examining how UVR affects developing marine organisms and cells in general. To model the protein-mediated cell cycle response to UV-irradiation, six batches of S. purpuratus embryos were exposed to UVR, monitored for delays in the first mitotic division and examined for global proteomic changes. Embryos from each batch were exposed to or protected from artificial UVR for 25 or 60 min. Embryos treated with …