Matching Items (24)

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The Development of a Plant-Expressed M2e-Based Universal Influenza Vaccine

Description

Influenza is a deadly disease for which effective vaccines are sorely lacking. This is largely due to the phenomena of antigenic shift and drift in the influenza virus's surface proteins, hemagglutinin (HA) and neuraminidase (NA). The ectodomain of the matrix

Influenza is a deadly disease for which effective vaccines are sorely lacking. This is largely due to the phenomena of antigenic shift and drift in the influenza virus's surface proteins, hemagglutinin (HA) and neuraminidase (NA). The ectodomain of the matrix 2 protein (M2e) of influenza A, however, has demonstrated high levels of conservation. On its own it is poorly immunogenic and offers little protection against influenza infections, but by combining it with a potent adjuvant, this limitation may be overcome. Recombinant immune complexes, or antigens fused to antibodies that have been engineered to form incredibly immunogenic complexes with one another, were previously shown to be useful, immunogenic platforms for the presentation of various antigens and could provide the boost in immunogenicity that M2e needs to become a powerful universal influenza A vaccine. In this thesis, genetic constructs containing geminiviral replication proteins and coding for a consensus sequence of dimeric M2e fused to antibodies featuring complimentary epitopes and epitope tags were generated and used to transform Agrobacterium tumefaciens. The transformed bacteria was then used to cause Nicotiana benthamiana to transiently express M2e-RICs at very high levels, with enough RICs being gathered to evaluate their potency in future mouse trials. Future directions and areas for further research are discussed.

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2018-05

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Is a putative plant-derived analog of the mammalian proline-rich attachment domain causing a human enzyme expressed in plants to undergo tetramerization?

Description

Variants of human butyrylcholinesterase (BChE) have been designed to have high cocaine hydrolytic activity. These variants have potential pharmacological applications toward treating cocaine overdose and addiction. These enzymes must be stable in the human body over fairly long periods of

Variants of human butyrylcholinesterase (BChE) have been designed to have high cocaine hydrolytic activity. These variants have potential pharmacological applications toward treating cocaine overdose and addiction. These enzymes must be stable in the human body over fairly long periods of time in order to be effective at treating cocaine addiction. Recombinantly expressed BChE, however, tends to be in monomer or dimer oligomeric forms, which are far less stable than the tetramer form of the enzyme. When BChE is transiently expressed in Nicotiana benthamiana, it is produced mainly as monomers and dimers. However, when the protein is expressed through stable transformation, it produces much greater proportions of tetramers. Tetramerization of WT human plasma derived BChE is facilitated by the binding of a proline rich peptide. In this thesis, I investigated if a putative plant-derived analog of the mammalian proline-rich attachment domain caused stably expressed cocaine hydrolase variants of human BChE to undergo tetramerization. I also examined if co-expression of peptides with known proline-rich attachment domains further shifted the monomer-tetramer ratio toward the tetramer.

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Date Created
2015-05

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Food Waste Fertilizer Efficacy

Description

As the world’s population exponentially grows, more food production is required. This increasing food production currently has led to the un-sustainable production of chemical fertilizers and resultant overuse. A more sustainable option to enhance food production could be the use

As the world’s population exponentially grows, more food production is required. This increasing food production currently has led to the un-sustainable production of chemical fertilizers and resultant overuse. A more sustainable option to enhance food production could be the use of fertilizer derived from food waste. To address this, we investigated the possibility of utilizing a fertilizer derived from food waste to grow hydroponic vegetables. Arugula (Eruca sativa) ‘Slow Bolt’ and lettuce (Lactuca sativa) ‘Cherokee’ and ‘Rex’ were cultivated using indoor deep-flow hydroponic systems at 23 ºC under a photosynthetic photon flux density of 170 µmol∙m−2∙s−1 with an 18-hour photoperiod. Plant nutrient solutions were provided by food waste fertilizer or commercial 15:5:20 NPK fertilizer at the identical electrical conductivity (EC) of 2.3 mS·cm–1. At the EC of 2.3 mS·cm–1, chemical fertilizer contained 150 ppm N, 50 ppm P, and 200 ppm K, while food waste fertilizer had 60 ppm N, 26 ppm P, and 119 ppm K. Four weeks after the nutrient treatments were implemented, compared to plants grown with chemical fertilizer, lettuce ‘Rex’ grown with food waste fertilizer had four less leaves, 27.1% shorter leaves, 68.2% and 23.1% less shoot and root fresh weight, respectively. Lettuce ‘Cherokee’ and arugula grown with food waste fertilizer followed a similar trend with fresh shoot weights that were 80.1% and 95.6% less compared to the chemical fertilizer, respectively. In general, the magnitude of reduction in the plant growth was greatest in arugula. These results suggest that both fertilizers were able to successfully grow lettuce and arugula, although the reduced plant growth with the food waste fertilizer in our study is likely from a lower concentration of nutrients when we considered EC as an indicator of nutrient concentration equivalency of the two fertilizer types.

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Date Created
2021-05

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Flora of Usery Mountain Regional Park and Pass Mountain region of Tonto National Forest, Arizona and distribution of Saguaro (Carnegniea gigantea) on Pass Mountain in southern Tonto National Forest

Description

This study was designed to produce a comprehensive flora of Usery Mountain Regional Park and Pass Mountain of the Tonto National Forest. A total of 168 vascular plant species representing 46 families and 127 genera were collected or documented at

This study was designed to produce a comprehensive flora of Usery Mountain Regional Park and Pass Mountain of the Tonto National Forest. A total of 168 vascular plant species representing 46 families and 127 genera were collected or documented at this study area. Sixteen species were not native to the flora of Arizona and represent 9.5% of the flora. Nevertheless, the study area does not appear to be significantly damaged or degraded in spite of its historical and current land use. The location and types of invasive species recorded in this study will assist with implementing preventative measures to prevent further spreading of certain species. The complete list of all vascular species recorded in this study will provide a valuable tool for land management decisions and future restoration projects that may occur at this area or similar sites and invasive species control. The distribution of the saguaro (Carnegiea gigantea) population on Pass Mountain was documented through the measurement of saguaros by random sampling. ArcGIS was used to generate 50 random points for sampling the saguaro population. Analysis to determine saguaro habitat preferences based on the parameters of aspect, slope and elevation was conducted through ArcGIS. The saguaro population of Pass Mountain significantly favored the southern aspects with the highest concentration occurring in the southwest aspects at an average density of 42.66 saguaros per hectare. The large numbers of saguaros recorded in the younger size classes suggests a growing populations.

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Date Created
2011

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Evaluation of Plant-based Viral Vectors for West Nile Virus Antibody Expression Levels

Description

Plant viral vectors have previously been used to produce high expression levels of antibodies and other proteins of interest. By utilizing a transformed Agrobacterium with the vector containing the protein of interest for infiltration, viral vectors can easily reach the

Plant viral vectors have previously been used to produce high expression levels of antibodies and other proteins of interest. By utilizing a transformed Agrobacterium with the vector containing the protein of interest for infiltration, viral vectors can easily reach the plant cells making it an effective form of transient protein expression. For this project two different plant viral vectors were compared; the geminiviral vector derived from Bean yellow dwarf virus (BeYDV) and the MagnICON vector system derived from Tobacco Mosaic Virus(TMV) and Potato Virus X(PVX). E16, an antibody against West Nile virus, has previously been expressed using both systems but expression levels between the systems were not directly compared. Agrobacterium tumefaciens EHA105 cells were transformed with both systems and expression levels of E16 were quantified using ELISAs. Results showed very low expression levels of E16 using the geminiviral vector indicating a need for further investigation into the clone used as previous studies reported much higher expression levels with the system.

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Date Created
2020-05

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Plant-derived Virus-like Particles and Recombinant Immune Complexes as Potential Components of a Future HIV Vaccine

Description

HIV continues to remain a global health issue, in particular in many low and middle-income countries. The World Health Organization (WHO) estimates that of the nearly 38 million HIV-1 positive individuals, 25% are unaware they are infected. Despite decades of

HIV continues to remain a global health issue, in particular in many low and middle-income countries. The World Health Organization (WHO) estimates that of the nearly 38 million HIV-1 positive individuals, 25% are unaware they are infected. Despite decades of research, a safe and effective preventative vaccine has yet to be produced. The HIV-1 envelope glycoprotein41 and the Gag structural protein have been identified to be particularly important in HIV-1 transcytosis and cytotoxic lymphocyte response, respectively. Enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of glycoprotein (dgp41) comprising the membrane proximal external region (MPER), transmembrane domain and cytoplasmic tail may present a unique and safe way of presenting these proteins in a state mimicking their natural formation. Another form of presenting the immunogenic glycoprotein41, particularly the MPER component, is by presenting it onto the N-terminal of an IgG molecule, thereby creating an IgG fusion molecule. In our lab, both VLPs and IgG fusion molecules are highly expressed and purified within GnGn Nicotiana benthamiana. The results indicated that these recombinant proteins can be assembled properly within plants and can elicit an immune response in mice. This provides a preliminary step in using such Gag/dpg41 VLPs and RIC as present a safe, effective, and inexpensive HIV vaccine.

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Date Created
2020-05

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The Supermarket Game: An Internet Teaching Tool Designed to Enhance Understanding of Economically Important Food Plants

Description

Plants are essential to human life. They release oxygen into the atmosphere for us to breathe. They also provide shelter, medicine, clothing, tools, and food. For many people, the food that is on their tables and in their supermarkets isn't

Plants are essential to human life. They release oxygen into the atmosphere for us to breathe. They also provide shelter, medicine, clothing, tools, and food. For many people, the food that is on their tables and in their supermarkets isn't given much thought. Where did it come from? What part of the plant is it? How does it relate to others in the plant kingdom? How do other cultures use this plant? The most many of us know about them is that they are at the supermarket when we need them for dinner (Nabhan, 2009) (Vileisis, 2008).

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Date Created
2012-12

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Advancing access to biodiversity data using the SALIX method and digital field guides

Description

The Arizona State University Herbarium began in 1896 when Professor Fredrick Irish collected the first recorded Arizona specimen for what was then called the Tempe Normal School - a Parkinsonia microphylla. Since then, the collection has grown to approximately 400,000

The Arizona State University Herbarium began in 1896 when Professor Fredrick Irish collected the first recorded Arizona specimen for what was then called the Tempe Normal School - a Parkinsonia microphylla. Since then, the collection has grown to approximately 400,000 specimens of vascular plants and lichens. The most recent project includes the digitization - both the imaging and databasing - of approximately 55,000 vascular plant specimens from Latin America. To accomplish this efficiently, possibilities in non-traditional methods, including both new and existing technologies, were explored. SALIX (semi-automatic label information extraction) was developed as the central tool to handle automatic parsing, along with BarcodeRenamer (BCR) to automate image file renaming by barcode. These two developments, combined with existing technologies, make up the SALIX Method. The SALIX Method provides a way to digitize herbarium specimens more efficiently than the traditional approach of entering data solely through keystroking. Using digital imaging, optical character recognition, and automatic parsing, I found that the SALIX Method processes data at an average rate that is 30% faster than typing. Data entry speed is dependent on user proficiency, label quality, and to a lesser degree, label length. This method is used to capture full specimen records, including close-up images where applicable. Access to biodiversity data is limited by the time and resources required to digitize, but I have found that it is possible to do so at a rate that is faster than typing. Finally, I experiment with the use of digital field guides in advancing access to biodiversity data, to stimulate public engagement in natural history collections.

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Date Created
2012

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The Phoenix Four Rivers Flora, Maricopa County, Arizona

Description

ABSTRACT The Phoenix Four Rivers Flora is an inventory of all the vascular plants growing along the Salt, Gila, New and Agua Fria Rivers, and their tributaries in the Phoenix Metropolitan Area during the years of the study (2009-2011).

ABSTRACT The Phoenix Four Rivers Flora is an inventory of all the vascular plants growing along the Salt, Gila, New and Agua Fria Rivers, and their tributaries in the Phoenix Metropolitan Area during the years of the study (2009-2011). This floristic inventory documents the plant species and habitats that exist currently in the project area, which has changed dramatically from previous times. The data gathered by the flora project thus not only documents how the current flora has been altered by urbanization, but also will provide a baseline for future ecological studies. The Phoenix Metropolitan Area is a large urbanized region in the Sonoran Desert of Central Arizona, and its rivers are important for the region for many uses including flood control, waste water management, recreation, and gravel mining. The flora of the rivers and tributaries within the project area is extremely diverse; the heterogeneity of the systems being caused by urbanization, stream modification for flood control, gravel mining, and escaped exotic species. Hydrological changes include increased runoff in some areas because of impermeable surfaces (e.g. paved streets) and decreased runoff in other areas due to flood retention basins. The landscaping trade has introduced exotic plant species that have escaped into urban washes and riparian areas. Many of these have established with native species to form novel plant associations.

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Date Created
2011

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Life of photosynthetic complexes in the cyanobacterium Synechocystis sp. PCC 6803

Description

The cyanobacterium Synechocystis sp. PCC 6803 performs oxygenic photosynthesis. Light energy conversion in photosynthesis takes place in photosystem I (PSI) and photosystem II (PSII) that contain chlorophyll, which absorbs light energy that is utilized as a driving force for photosynthesis.

The cyanobacterium Synechocystis sp. PCC 6803 performs oxygenic photosynthesis. Light energy conversion in photosynthesis takes place in photosystem I (PSI) and photosystem II (PSII) that contain chlorophyll, which absorbs light energy that is utilized as a driving force for photosynthesis. However, excess light energy may lead to formation of reactive oxygen species that cause damage to photosynthetic complexes, which subsequently need repair or replacement. To gain insight in the degradation/biogenesis dynamics of the photosystems, the lifetimes of photosynthetic proteins and chlorophyll were determined by a combined stable-isotope (15N) and mass spectrometry method. The lifetimes of PSII and PSI proteins ranged from 1-33 and 30-75 hours, respectively. Interestingly, chlorophyll had longer lifetimes than the chlorophyll-binding proteins in these photosystems. Therefore, photosynthetic proteins turn over and are replaced independently from each other, and chlorophyll is recycled from the damaged chlorophyll-binding proteins. In Synechocystis, there are five small Cab-like proteins (SCPs: ScpA-E) that share chlorophyll a/b-binding motifs with LHC proteins in plants. SCPs appear to transiently bind chlorophyll and to regulate chlorophyll biosynthesis. In this study, the association of ScpB, ScpC, and ScpD with damaged and repaired PSII was demonstrated. Moreover, in a mutant lacking SCPs, most PSII protein lifetimes were unaffected but the lifetime of chlorophyll was decreased, and one of the nascent PSII complexes was missing. SCPs appear to bind PSII chlorophyll while PSII is repaired, and SCPs stabilize nascent PSII complexes. Furthermore, aminolevulinic acid biosynthesis, an early step of chlorophyll biosynthesis, was impaired in the absence of SCPs, so that the amount of chlorophyll in the cells was reduced. Finally, a deletion mutation was introduced into the sll1906 gene, encoding a member of the putative bacteriochlorophyll delivery (BCD) protein family. The Sll1906 sequence contains possible chlorophyll-binding sites, and its homolog in purple bacteria functions in proper assembly of light-harvesting complexes. However, the sll1906 deletion did not affect chlorophyll degradation/biosynthesis and photosystem assembly. Other (parallel) pathways may exist that may fully compensate for the lack of Sll1906. This study has highlighted the dynamics of photosynthetic complexes in their biogenesis and turnover and the coordination between synthesis of chlorophyll and photosynthetic proteins.

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Date Created
2011