Biochemistry Commons^™

Study Of Protein Extraction Techniques For M. Foliorum Phages, Lola Sibaud, Anna Elgersma

Student Academic Conference

Characterizing phages is becoming increasingly important in biochemistry and biotechnology due to their potential to treat bacterial infections without need for antibiotics. Last year, students attempted to isolate proteins from Microbacterium foliorum infecting phages Babydotz and Rosadiaz. Proteins were successfully extracted; however, after further analysis it was shown that the obtained proteins belonged to the host bacteria and not the phages. The purpose of this study was to find a method to separate phage from host bacterial debris to successfully isolate phage proteins and analyze them later. Isolating phage proteins could help us characterize more phages and know more about …

Endogenous Cftr Expression In Human Epithelial Cell Lines, Zithlaly Amezquita

Undergraduate Research Conference

CFTR (Cystic Fibrosis Transmembrane-conductance Regulator) is a plasma membrane protein that functions as a chloride ion channel on many epithelial cells. There are over 1000 mutations that affect the function of this protein; however, the most common mutation is DF508. CFTR mutations target many secretory organs like pancreas and lungs causing the genetic disease cystic fibrosis (CF), common among Caucasians of northern European origin. Detection of endogenous CFTR expression is possible by a difficult and costly immunoprecipitation method but not by the most used western blotting method. Therefore, the goal of this project was to detect the endogenous CFTR expression …

Investigating Stop Codon Readthrough In Saccharomyces Cerevisiae, Amy Kwon

Undergraduate Student Research Internships Conference

Stop codon readthrough occurs via genetic and epigenetic mechanisms, resulting in a longer polypeptide chain at the C-terminus. Although these readthroughs may seem like an error made by translational mechanisms, evidence from yeast suggests that stop codon readthrough has an impact on various cellular processes. Readthrough has the potential to create genetic diversity, similar to RNA splicing, mRNA editing, and protein modification. The diversity created by translational readthrough may result in a beneficial change in phenotype, and thus have a role in evolution and adaptability. Translational readthrough is observed in organisms such as yeast and E. coli but may also …

Niche Partitioning And Utilization Of Different Nitrogen Sources By Marine Cyanobacteria Synechococcus, Angel Bui, Anne W. Thompson

McNair Symposium

The Cyanobacterium Synechococcus plays a major role in the ocean’s biochemical processes and is responsible for a significant amount of primary production, especially around coastal areas. Synechococcus has a wide geographical distribution that includes both polar and high-nutrient waters. Within the genus, there are defined subpopulations that are ecologically distinct that allow them to niche partition the dynamic oceans. To further explore niche partitioning of Synechococcus, this project combines a bioinformatic and culture-based approach. I examined data along the North Pacific Subtropical Front (NSPF) to analyze the community structure. This analysis demonstrated that a particular ecotype identified as clade II …

Cd40l And Tnf-Alpha Levels After Traumatic Injury, Queen Revollido, Martin Schreiber

McNair Symposium

The biomarkers CD40 ligand and TNF-alpha are proteins that exhibit proinflammatory and prothrombotic effects. A small clinical study has shown that soluble CD40 ligand may have a role in early coagulopathy and inflammatory complications in severely injured patients. It is known that a CD40 ligand increases the production of cytokines such as TNF-alpha, thus, the current study aims to investigate the kinetics of these biomarkers and the extent of upregulation after traumatic injury. Frozen samples collected from baseline, 8, 24, and 48 hours after admission will be used. CD40 ligand and TNF-alpha levels will be quantified using Luminex that will …

Analysis Of Botulinum Toxin A And Interacting Proteins In Skeletal Muscle Cells: An Investigation Into The Mechanisms Of Botulinum Toxin A As A Treatment For Chronic Exertional Compartment Syndrome, Lauren Kee

Pence-Boyce STEM Student Scholarship

Background: Chronic exertional compartment syndrome (CECS) is a condition in which muscle tissue expands against the surrounding fascia during activity and is compressed along with the nerves and blood vessels within the muscle compartment, leading to abnormally high intracompartmental pressure (ICP) and debilitating pain. Treatment typically includes fasciotomy, which results in significant levels of CECS recurrence; however, botulinum toxin A (BoNT-A) injection has recently been seen to decrease both ICP and pain through an unknown mechanism with little to no recurrence.

Methods: In this study, cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), prostaglandin E₂ receptor 4 (EP4), endothelial nitric oxide synthase (eNOS), …

Morph- And Sex-Specific Differences In Corticosterone Of The Arizona Tiger Salamander (Ambystoma Mavortium Nebulosum), Megan Zerger

Scholars Week

Life history morph, sex, and body condition are traits that may influence stress within salamander populations because of differences in physiology and environmental conditions. Given widespread declines and the effects chronic stress can have on amphibian health, it is important to understand within-population drivers of stress and how population level variation may influence population viability. Thus, the objective of our study was to assess how corticosterone varies within the Arizona tiger salamander (Ambystoma mavortium nebulosum) population at the Mexican Cut Nature Preserve. We used a non-invasive skin swabbing method to collect baseline and elevated corticosterone from paedomorph (aquatic …

Insights Into Viral Genome Function Through Comparative Structural Analysis, Lydia Phillips

UNO Student Research and Creative Activity Fair

Enteroviruses are single stranded RNA viruses which have caused many public health concerns, particularly in children. These viruses are responsible for polio, hand, foot, and mouth disease, many polio-like neurological diseases, and the common cold. The enterovirus called Coxsackievirus B3 (CVB3), a close relative to poliovirus, has been shown responsible for severe human disease, including pancreatitis, myocarditis, and type 1 diabetes. A unique advantage of studying CVB3 is the existence of a naturally occurring strain (CVB3/GA) which displays no known pathogenicity. Earlier comparative genomic studies have shown that the primary difference between avirulent CVB3/GA and the virulent strains of CVB3 …

Improvement In 14-3-3 Binding Site Prediction, Katherine K. Mccormack

ScholarsArchive Data

The 14-3-3 family of phospho-binding proteins regulate a variety of major cellular processes through interaction with a network of dynamic proteins. Deregulation of the 14-3-3 interaction network contributes to a variety of degenerative disorders and cancers. Our lab focuses on identifying novel 14-3-3 interactions and understanding how 14-3-3 binding regulates protein function. A major gap in this process is that identifying the phospho-site where 14-3-3 docks on a given protein is time- and resource-consuming. Prediction algorithms have been developed to predict canonical 14-3-3 binding sites, however, there are many non-canonical sites that existing software is unable to predict. To fill …

Progress Presentation On The Proposed Function Of Protein 4diu Through Bioinformatics Analyses, Melissa Hoff

Biochemistry and Molecular Biology Presentations

The BASIL project is a scientific inquiry project that aims to determine the function of a protein from its structure. Biochemistry students completed wet lab work, while structural biology students worked in silico. The purpose of this study was to determine the function of the protein with PDB ID: 4DIU. Bioinformatics analyses were completed using several databases: BLAST, Dali, Pfam, and Moltimate, and imaging of the protein was completed using PyMOL software. The BLAST database aligned the primary sequence with known protein sequences. The Dali database compared protein structure with known structures through global alignment. The Pfam database matched our …

The Classification Of An Unknown Protein 3h04, Britney Dyszel

Biochemistry and Molecular Biology Presentations

An unclassified protein in the Protein Data Bank, 3H04, was selected as a protein of interest. Research was performed in silico to gain key information on the structure, sequence, and homology of protein 3H04. This research was guided using the BASIL Project’s protocols on in silico research. Several databases were utilized to study protein 3H04. Based on the data gathered, unclassified protein 3H04 is an aminopeptidase that cleaves prolyl-dipeptidyl peptide bonds through alpha beta hydrolase function.

Opening Of An Epoxide Ring Using Azide To Form A Triazole, Emily Hufnagel, Jeffrey A. Hansen

Annual Student Research Poster Session

No abstract provided.

Synthesis Of Ether Alcohols With Varying Catalysts, Hannah Grady, Frida Li, Jeffrey A. Hansen

Annual Student Research Poster Session

No abstract provided.

Targeted Therapy For The Future: The Use Of Novel Antimicrobial Peptides Against P. Aeurginosa, Matthew Froid

UNO Student Research and Creative Activity Fair

Pathogenic bacteria, such as the gram-negative bacterium Pseudomonas aeruginosa, are becoming resistant to our current arsenal of antibiotics at an alarming rate. P. aeruginosa is a leading cause of nosocomial acquired infections and is a primary co-morbidity in patients with compromised immune systems. One potential source of new antibiotic agents is antimicrobial peptides. Antimicrobial peptides (AMPs) are small proteins, and some have shown a high degree of efficacy and broad-spectrum activity against both Gram-positive and Gram-negative bacteria. An experimental AMP that has been developed by Dr. Wang at UNMC, DASamp2, has shown to be effective against virulent bacteria, including P. …

Large Scale Dynamical Model Of Macrophage/Hiv Interactions, Matthew M. Froid

UNO Student Research and Creative Activity Fair

Properties emerge from the dynamics of large-scale molecular networks that are not discernible at the individual gene or protein level. Mathematical models - such as probabilistic Boolean networks - of molecular systems offer a deeper insight into how these emergent properties arise. Here, we introduce a non-linear, deterministic Boolean model of protein, gene, and chemical interactions in human macrophage cells during HIV infection. Our model is composed of 713 nodes with 1583 interactions between nodes and is responsive to 38 different inputs including signaling molecules, bacteria, viruses, and HIV viral particles. Additionally, the model accurately simulates the dynamics of over …

A Crispr Platform For Rapid And Inducible Genome Editing In Human Non-Small Cell Lung Cancer Cells, Lloyd Bartley

Posters-at-the-Capitol

Non-small cell lung cancer (NSCLC) accounts for about 85% of lung cancer, which is the leading cause of cancer death in the world. High mortality rate associated with NSCLC is partially attributed to the limited understanding of NSCLC as well as ineffective therapeutic treatments. The initiation and progression of NSCLC involves genetic changes leading to alterations in the control of tissue development and homeostatic maintenance. Better knowledge about these genetic abnormalities is imperative for developing new chemotherapeutic drugs for NSCLC. Recent research demonstrates that the expression of paraoxonase 2 (PON2), a lactonase/arylesterase with anti-oxidant properties, are markedly enhanced in cancer …

Synthesis And Incorporation Of 1,2-Alkanolamine-Functionalized Lysine As A Non-Canonical Amino Acid Into Gfp, Chesley M. Rowlett

Scholar Week 2016 - present

Synthesis of specific post translational modifications in proteins can be difficult but achievable via genetic code expansion techniques. An attempt has been made to synthesize and incorporate D-cThrK into green fluorescent protein (GFP) at an amber mutation site in Escherichia coli via the coordination of pyrrolysyl- tRNA synthetase and its cognate tRNApyl. The incorporation of this non-canonical amino acid and potential chemical transformations following it allow the synthesis of proteins with post translational lysine modifications, making a variety of basic and biotechnological applications available.

Elemental Analysis Of Beer Manufactured In Virginia Satellite Breweries, Jennifer Wiliams

Student Scholar Showcase

Financial incentives from the state and wider distribution potential have lured many west coast brewers to open satellite breweries in Virginia. Quality control is crucial for any brewery and especially important in satellite breweries that need to imitate the standard set by the parent brewery. Zinc, magnesium, iron, sodium manganese, and calcium are trace elements found in wort and are essential for enzymes that catalyze fermentation reactions. These elements protect against cell stress, play a crucial role in flocculation, and can affect the flavoring and coloration of beer. Barley, water, and hops are the sources of these elements in beer. …

Studies Of Amino Acid Mutations In Drug Resistance Of The Smo Protein, Eunice Wintona

UNO Student Research and Creative Activity Fair

Smoothened receptor (SMO) is a protein that in humans, is encoded by the SMO gene. A systemic mutation in its binding pocket helps predict the sensitivity of mutant proteins to different drugs. Known as a GPCR-like receptor, it is a component of the hedgehog signaling pathway; a pathway involved in body patterning and the regulation of adult stem cells. An uncontrolled or inappropriate activation of the Hedgehog pathway drives tumor progression in cancers and a number of birth defects. To achieve these goals, the molecular modeling software MOE was used to build small molecules and drug molecules like Vismodegib and …

Fatty Acid Recovery And Identification In Mars Analogue Soil Samples, Kimberly E. Lykens

STAR Program Research Presentations

A primary goal of NASA’s Mars 2020 mission is to gather and store samples of Mars soil that could possibly be returned to Earth in a future mission for investigations into past or present life. In years leading up to a potential sample return mission on Mars and potentially other worlds, samples taken from Chile’s Atacama Desert, the most arid, biologically limited desert in the world, are valuable in developing a capacity for biosignature detection, specifically when exploring fatty acid abundance. Eighteen samples were collected from two sites in the Atacama characterized by biological soil crusts (BSC). BSCs are areas …

Repsa-Directed Identification Of Dna-Binding Specificity For Orphan Transcription Factors, Kamir Hiam

Symposium of Student Scholars

The function of many genes and the biological roles of their encoded products are still not well characterized. Given the sequence-specific DNA-binding properties of transcription factor proteins, it is possible to purify them, identify the responsible polypeptide(s), determine their consensus binding sequences, and identify their genomic binding sites. Thus, one can go from cellular extract to proposed biological regulatory roles in relatively short order. Our goal is to identify and characterize orphan DNA-binding proteins in the model organism E. coli K12 using the novel combinatorial technique, REPSA (Restriction Endonuclease Protection Selection Amplification), as well as further develop the REPSA to …