Medicine and Health Sciences Commons^™

Glutaredoxin 2 (Grx2) Knockout Increases Sensitivity To Oxidative Stress In Mouse Lens Epithelial Cells, Hongli Wu, Liren Lin, Frank Giblin, Ye-Sheh Ho, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

Glutaredoxin belongs to the oxidoreductase family with cytosolic glutaredoxin 1 (Grx1) and mitochondrial gluraredoxin 2 (Grx2) isoforms. Of the two isozymes, the function of Grx2 is not well understood. This paper studied the effect of Grx2 deletion on cellular function using primary lens epithelial cell cultures isolated from Grx2 gene knockout (KO) and wild type (WT) mice. We found that both cell types showed similar growth patterns and morphology, and comparable mitochondrial glutathione pool and complex I activity. Cells with deleted Grx2 did not show affected Grx1 or thioredoxin (Trx) expression but exhibited high sensitivity to oxidative stress. Under treatment …

Spectrum Of Acute Clinical Characteristics Of Diagnosed Concussions In College Athletes Wearing Instrumented Helmets, Ann-Christine Duhaime,, Jonathan G. Beckwith, Arthur C. Maerlender, Thomas W. Mcallister, Joseph J. Crisco, Stefan M. Duma, P. Gunnar Brolinson, Steven Rowson, Laura A. Flashman, Jeffrey J. Chu, Richard M. Greenwald

Department of Psychology: Faculty Publications

Object. Concussive head injuries have received much attention in the medical and public arenas, as concerns have been raised about the potential shortand long-term consequences of injuries sustained in sports and other activities. While many student athletes have required evaluation after concussion, the exact definition of concussion has varied among disciplines and over time. The authors used data gathered as part of a multiinstitutional longitudinal study of the biomechanics of head impacts in helmeted collegiate athletes to characterize what signs, symptoms, and clinical histories were used to designate players as having sustained concussions.

Methods. Players on 3 college …

Structure-Function Analysis Of A Protein Encoded By The Bhv-1 Latency Related Gene, Devis Sinani

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Bovine herpes virus 1 (BHV-1) is a significant viral pathogen in cattle that induces a myriad of clinical symptoms. These symptoms include: conjunctivitis, upper respiratory tract infections, genital disorders, and abortions. BHV-1 infection can also lead to transient immune-suppression, which predisposes cattle to secondary bacterial infection leading to life-threatening pneumonia referred to as bovine respiratory disease (BRD). Following acute infection, BHV-1 establishes latency in sensory neurons within trigeminal ganglia. Reactivation of the virus can occur periodically, resulting in virus transmission. The latency-related (LR) RNA is the only abundantly expressed transcript in latently infected sensory neurons and it encodes several proteins, …

Measuring Parvalbumin Levels In Fish Muscle Tissue: Relevance Of Muscle Locations And Storage Conditions, Poi-Wah Lee, Julie A. Nordlee, Stef J. Koppelman, Joseph L. Baumert, Steve L. Taylor

Department of Food Science and Technology: Faculty Publications

Fish is an allergenic food capable of provoking severe anaphylactic reactions. Parvalbumin is the major allergen identified in fish and frog muscles. Antibodies against fish and frog parvalbumin have been used to quantify parvalbumin levels from fish. However, these antibodies react variably with parvalbumin from different fish species. Several factors might be responsible for this variation including instability of parvalbumin in fish muscle as a result of frozen storage and differential parvalbumin expression in muscles from various locations within the whole fish. We aimed to investigate whether these factors contribute to the previously observed variable immunoreactivity of the anti-parvalbumin antibodies. …

School Food Environment The Frontline For Childhood Obesity Prevention: A Mixed-Method Study Of Nutritional Competencies And Skills Of School Nutriiton Professionals In Nebraska, Zainab Rida

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

The purpose of this statewide study was to address the nutrition knowledge, attitudes, and perceptions of school foodservice personnel in Nebraska regarding offering/serving healthy school meals. Moreover, this study identified some potential barriers and avenues of action for offering/serving healthy school meals. Data obtained from this study indicates that there is a strong correlation (r= .103, p r .237, p< .01) between the foodservice staff’s self-efficacy and their practices of offering/serving healthy school meals. Fortunately, the relationship between foodservice staff practices of offering/serving healthy school meals and their self-efficacy was positive and significantly predicted practices scores, β =.237, P< 0.01. The finding of the present study also identifies many barriers including lack of time and support that face the foodservice personnel in offering/serving healthy school meals. The findings suggested that there is an urgent need of a full school approach to promote and encourage healthy eating habits among students. Future research is needed to evaluate school wellness policies regarding healthy eating practices in schools. Moreover, establish partnerships with communities and universities for intervention that target students and their parents.

Advisor: Wanda Koszewski

Medical Nutrition Therapy Provided To Adult Hematopoietic Stem Cell Transplantation Patients, Mary G. Habschmidt, Cheryl A. Bacon, Mary B. Gregoire, Heather E. Rasmussen

Department of Nutrition and Health Sciences: Faculty Publications

This study explored the current medical nutrition therapy (MNT) provided to adult patients undergoing hematopoietic stem cell transplantation (HSCT) and examined the current and desired role of registered dietitians (RDs) in providing MNT. A total of 60 RDs (57% response rate) responded to an electronic questionnaire. Descriptive statistics and χ² analyses (SPSS, version 18) were used. Results revealed the primary form of diet was oral, and for patients on nutrition support, parenteral nutrition (PN) was used more frequently (16%–31%) than enteral nutrition (EN) (5%–9%; P ≤ .05). Nutrition support decisions were based on patients’ individualized needs rather than established …

Ultraviolet Radiation–Induced Cataract In Mice: The Effect Of Age And The Potential Biochemical Mechanism, Jie Zhang, Hong Yan, Stefan Lofgren, Xiaoli Tian, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

PURPOSE. To study the effect of age on the morphologic and biochemical alterations induced by in vivo exposure of ultraviolet radiation (UV).

METHODS. Young and old C57BL/6 mice were exposed to broadband UVBþUVA and euthanized after 2 days. Another batch of UV-exposed young mice was monitored for changes after 1, 2, 4, and 8 days. Age-matched nonexposed mice served as controls. Lens changes were documented in vivo by slit-lamp biomicroscopy and dark field microscopy photographs ex vivo. Lens homogenates were analyzed for glutathione (GSH) level, and the activities of thioredoxin (Trx), thioltransferase (TTase), and glyceraldehyde-3-phosphate dehydrogenase (G3PD). Glutathionylated lens proteins …

Parvalbumin In Fish Skin–Derived Gelatin: Is There A Risk For Fish Allergic Consumers?, S. J. Koppelman, J. A. Nordlee, P.-W. Lee, R. P. Happe, M. Hessing, R. Norland, T. Manning, R. Deschene, G. A. H. De Jong, S. L. Taylor

Department of Food Science and Technology: Faculty Publications

The major allergen parvalbumin was purified from cod muscle tissues, and polyclonal antibodies were raised toward it. The antibodies were tested for specificity, and an enzyme-linked immunosorbent assay (ELISA) was developed using these antibodies. The ELISA was applied to measure parvalbumin in cod skin, the starting material for fish gelatin made from deep sea, wild fish. The ELISA was sufficiently sensitive (LLOQ = 0.8 ng ml^–1 in extracts, corresponding to 0.02 μg of parvalbumin per g of tissue) and did not cross-react with common food constituents. Fish gelatin, wine, and beer, matrices for the potential use of this ELISA, …

Mutations In Dmrt3 Affect Locomotion In Horses And Spinal Circuit Function In Mice, Lisa S. Andersson, Martin Larhammar, Fatima Memic, Hanna Wootz, Doreen Schwochow, Carl-Johan Rubin, Kalicharan Patra, Thorvaldur Arnason, Lisbeth Wellbring, Göran Hjälm, Freyja Imsland, Jessica Lynn Petersen, Molly E. Mccue, James R. Mickelson, Gus Cothran, Nadav Ahituv, Lars Roepstorff, Sofia Mikko, Anna Vallstedt, Gabriella Lindgren, Leif Andersson, Klas Kullander

Department of Animal Science: Faculty Publications

Locomotion in mammals relies on a central pattern-generating circuitry of spinal interneurons established during development that coordinates limb movement. These networks produce left–right alternation of limbs as well as coordinated activation of flexor and extensor muscles. Here we show that a premature stop codon in the DMRT3 gene has a major effect on the pattern of locomotion in horses. The mutation is permissive for the ability to perform alternate gaits and has a favorable effect on harness racing performance. Examination of wild-type and Dmrt3-null mice demonstrates that Dmrt3 is expressed in the dI6 subdivision of spinal cord neurons, takes …

A Pilot Study: The Use Of A Survey To Assess The Food Knowledge Of Nutrition Students At Various Levels Of Nutrition Education, Chante Chambers

College of Education and Human Sciences: Dissertations, Theses, and Student Research

A working definition of a concept known as ‘food literacy’ encompasses using basic food preparation knowledge that has been learned, understood, and practiced to make better food decisions. To advance these skills for client service, a post-secondary nutrition program would need to include objectives that allow application of knowledge. For this reason, the purpose of this study was to determine the difference in knowledge and application among students in 100, 200 and 400 level college nutrition course(s). A survey was developed to measure the food knowledge of these college students as they prepare for careers as health care professionals. The …

Melanopsin Mediates Retrograde Visual Signaling In The Retina, Dao-Qi Zhang, Michael A. Belenky, Patricia J. Sollars, Gary E. Pickard, Douglas G. Mcmahon

School of Veterinary and Biomedical Sciences: Faculty Publications

The canonical flow of visual signals proceeds from outer to inner retina (photoreceptors→bipolar cells→ganglion cells). However, melanopsin-expressing ganglion cells are photosensitive and functional sustained light signaling to retinal dopaminergic interneurons persists in the absence of rods and cones. Here we show that the sustained-type light response of retinal dopamine neurons requires melanopsin and that the response is mediated by AMPA-type glutamate receptors, defining a retrograde retinal visual signaling pathway that fully reverses the usual flow of light signals in retinal circuits.

Evaluation Of A Social Marketing Campaign: 4 Day Throw Away, Katie J. James

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Children are at a heightened risk for foodborne illness. Handling leftover food safely was identified as a concern from focus groups with the main food preparer in families with young children in two Midwestern states. To design, implement, and evaluate a theory-based food safety education intervention to raise awareness of the USDA guideline for handling leftover food. The food safety social marketing campaign “4 Day Throw Away” launched in towns throughout Nebraska and Iowa. The campaign included both traditional and social media methods to relay the message of discarding leftovers after four days. A total of 600 guardians of young …

Sleep Hygiene And Problem Behaviors In Snoring And Non- Snoring School-Age Children, Lisa A. Witcher, David Gozal, Dennis L. Molfese, Scott M. Salathe, Karen Spruyt, Valerie Mclaughlin Crabtree

Center for Brain, Biology, and Behavior: Faculty and Staff Publications

Objectives—The effects of sleep-disordered breathing, sleep restriction, dyssomnias, and parasomnias on daytime behavior in children have been previously assessed. However, the potential relationship(s) between sleep hygiene and children’s daytime behavior remain to be explored. The primary goal of this study was to investigate the relationship between sleep hygiene and problematic behaviors in non-snoring and habitually snoring children.

Methods—Parents of 100 5- to 8-year-old children who were reported to snore “frequently” to “almost always,” and of 71 age-, gender-, and ethnicity-matched children who were reported to never snore participated in this study. As part of a larger, ongoing study, …

Impact Of The Home Food Environment On Dietary Intake, Obesity And Cardiovascular Health Of U.S. Children And Adolescents, Aged 6-19, Melissa A. Masters

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Youth obesity is an ongoing problem in the United States. Obese children and adolescents are likely to be obese as adults and have an increased risk of developing chronic diseases, including coronary heart disease, earlier in life. The multifactorial nature of obesity continues to challenge researchers and health professionals to determine methods for preventing and reducing childhood obesity. Research has suggested that obesity is a normal response to an “obesigenic” environment. Emerging as one of the most influential environments in obesity and behavior development is the home food environment. However, little is understood about the role of the home food …

Dietitian’S Problem Solving Knowledge To Promote And Support Breastfeeding, Cassandra L. Dytrych

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

As a result of ongoing efforts in the U.S. to promote breastfeeding as the normal way to nourish an infant, breastfeeding initiation rates are higher than they have been in decades. As the experts on food and nutrition through the lifecycle, registered dietitians (RDs) should be knowledgeable to promote and support breastfeeding. It is not well known if RDs have this knowledge. The purpose of this study was to determine the current knowledge of RDs about breastfeeding, the types of professional opportunities RDs have to promote and support breastfeeding, and the level of education RDs have to deal with breastfeeding …

A Century And A Half Of Research On The Stable Fly, Stomoxys Calcitrans (L.) (Diptera: Muscidae), 1862-2011: An Annotated Bibliography, K. M. Kneeland, Steven R. Skoda, Jerome A. Hogsette, A. Y. Li, Jaime Molina-Ochoa, Kimberly Hutchinson Lohmeyer, John E. Foster

Department of Entomology: Faculty Publications

The stable fly, Stomoxys calcitrans, is a cosmopolitan pest of livestock, wild animals, pets, and humans. It is a primary pest of cattle in the United States, estimated to cause more than $1 billion in economic losses annually. It also causes dissension at the rural-urban interface and is a problem in recreation areas such as Florida beaches and the Great Lakes. Due to its pestiferous nature and painful bite, methods to control stable flies have been investigated for over a century. A large amount of research has been reported on stable fly biology, ecology, genetics, physiology, and vector competence. …

Autism Spectrum Disorders In Hispanics And Non-Hispanics, Virginia Chaidez, Robin L. Hansen, Irva Hertz-Picciotto

Department of Nutrition and Health Sciences: Faculty Publications

Objectives: To compare differences in autism between Hispanics and non-Hispanics. We also examined the relationship between multiple language exposure and language function and scores of children. Methods: The Childhood Autism Risks from Genetics and the Environment (CHARGE) study is an ongoing population-based case-control study with children sampled (n = 1061) from three strata: those with autism (AU) or autism spectrum disorder (ASD); developmental delay (DD); or the general population (GP). Results: Non-Hispanic cases demonstrated higher cognitive composite scores for the Mullen Scales of Early Learning (MSEL). There were significant associations between multiple language exposure and MSEL subscales for receptive …

Cell-Mediated Immunity In Porcine Reproductive And Respiratory Syndrome Virus, Rajeshwari Parida

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus is a significant swine pathogen which exhibits considerable sequence diversity. In an attempt to identify highly conserved T-cell epitopes contained in proteins of this virus, we examined heptadecamer peptides spanning the sequence of the PRRSV nonstructural proteins 9, 10 and 11, all of them are highly conserved, for their ability to elicit a recall proliferative and interferon-gamma response in peripheral blood mononuclear cells obtained from pigs immunized against the type-II PRRSV strain FL-12. These studies led to the identification of seven peptides, two from each NSP 9 and NSP 10 and, three from NSP …

Breakfast Consumption Habits Of Youth Living In Low Income Neighborhoods In A Large Three County Metropolitan Area, Elijah Dacy

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Purpose: The purpose of this study was to identify youth attitudes, frequency of, and barriers for breakfast consumption and the use of available nutrition programs by youth living in poverty or food insecure households in a large three county metropolitan area. Methods: A phone survey was developed and tested in order to collect information on breakfast consumption habits and attitudes of children, targeted as food insecure. There were no incentives for persons to participate in the survey and all information gathered was kept in a secure place at the Center for Human Nutrition. Participants who were allowed to …

Mixed Methods Analysis Of School Wellness Programs In Nebraska And Indiana: A Descriptive Study, Jessica A. Robinson

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Schools have become one of the major fronts that the battle against childhood obesity is being fought. In 2004 the Child Nutrition Act passed by Congress required all school districts receiving funds for the National School Lunch Program to create a wellness policy. Since then there have been many research projects conducted to learn more about these policies, and their implementation as programs in the schools. Thus far, no such research has been done in Nebraska or Indiana. The purpose of this descriptive study is to describe how schools in Nebraska and Indiana are implementing wellness programs in their schools. …

Flowcell Systems For Single Molecule Detection, John G. K. Williams, Gregory R. Bashford, Jiyan Chen, Dan Draney, Nara Narayanan, Bambi L. Reynolds, Pamela Sheaff

Biomedical Imaging and Biosignal Analysis Laboratory

The present invention provides compounds, methods and systems for sequencing nucleic acid using single molecule detection. Using labeled NPs that exhibit charge-switching behavior, single-molecule DNA sequencing in a microchannel sorting system is realized. In operation, sequencing products are detected enabling real-time sequencing as successive detectable moieties flow through a detection channel. By electrically sorting charged molecules, the cleaved product molecules are detected in isolation without interference from unincorporated NPs and without illuminating the polymerase-DNA complex.

Characterization Of Species C Human Adenovirus Serotype 6 (Ad6), Eric A. Weaver, Reeti Khare, Mathew L. Hillestad, Donna Palmer, Philip Ng, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Adenovirus serotype (Ad5) is the most studied Ad. Ad1, 2, and 6 are also members of species C Ad and are presumed to have biologies similar to Ad5. In this work, we have compared the ability of Ad1, 2, 5, and 6 to infect liver and muscle after intravenous and intramuscular injection. We found that Ad6 was surprisingly the most potent at liver gene delivery and that Ad1 and Ad2 were markedly weaker than Ad5 and 6. To understand these differences, we sequenced the Ad6 genome. This revealed that the Ad6 fiber protein is surprisingly three shaft repeats shorter than …

Heme Oxygenase-1 Regulates The Immune Response To Influenza Virus Infection And Vaccination In Aged Mice, Nathan W. Cummins, Eric A. Weaver, Shannon M. May, Anthony J. Croatt, Oded Foreman, Richard B. Kennedy, Gregory A. Poland, Michael A. Barry, Karl A. Nath, Andrew D. Badley

Nebraska Center for Virology: Faculty Publications

Underlying mechanisms of individual variation in severity of influenza infection and response to vaccination are poorly understood. We investigated the effect of reduced heme oxygenase-1 (HO-1) expression on vaccine response and outcome of influenza infection. HO-1-deficient and wild-type (WT) mice (kingdom, Animalia; phylum, Chordata; genus/species, Mus musculus) were infected with influenza virus A/PR/8/34 with or without prior vaccination with an adenoviral-based influenza vaccine. A genome-wide association study evaluated the expression of single-nucleotide polymorphisms (SNPs) in the HO-1 gene and the response to influenza vaccination in healthy humans. HO-1-deficient mice had decreased survival after influenza infection compared to WT mice (median …

Identification And Analysis Of The Ige Binding By Parvalbumin And Other Potential Allergens In Different Fish And Frog Species, P. Lee, J. A. Nordlee, S. J. Koppelman, J. L. Baumert, S. L. Taylor

Department of Food Science and Technology: Faculty Publications

Rationale: Serological cross-reactivity to different fish and frog species is common among fish-allergic individuals.We examined the intra- and inter-individual diversity in IgE responses of fish-allergic subjects to various fish and frog species and identified novel allergens besides parvalbumin.

Methods: Sera from 38 subjects with a clinical history of fish allergy were analyzed for IgE-binding profiles to crude extracts of 26 raw fish and frog species, and purified cod and carp parvalbumin using IgE-immunoblotting. Sera of 7 subjects showing similar IgE-binding profiles in the IgEimmmunoblotting were pooled to identify potential allergens in pilchard, herring, cod, cusk, and rainbow trout using two-dimensional …

Divergent Functions Of The Myotubularin (Mtm) Homologs Atmtm1 And Atmtm2 In Arabidopsis Thaliana: Evolution Of The Plant Mtm Family, Yong Ding, Ivan Ndamukong, Yang Zhao, Yuannan Xia, Jean-Jack Riethoven, David R. Jones, Nullin Divecha, Zoya Avramova

School of Biological Sciences: Faculty Publications

Myotubularin and myotubularin-related proteins are evolutionarily conserved in eukaryotes. Defects in their function result in muscular dystrophy, neuronal diseases, and leukemia in humans. In contrast to the animal lineage, where genes encoding both active and inactive myotubularins (phosphoinositide 3-phosphatases) have appeared and proliferated in the basal metazoan group, myotubularin genes are not found in the unicellular relatives of green plants. However, they are present in land plants encoding proteins highly similar to the active metazoan enzymes. Despite their remarkable structural conservation, plant and animal myotubularins have significantly diverged in their functions. While loss of myotubularin function causes severe disease phenotypes …

Copper–Zinc Superoxide Dismutase-Deficient Mice Show Increased Susceptibility To Experimental Autoimmune Encephalomyelitis Induced With Myelin Oligodendrocyte Glycoprotein 35–55, Chandirasegara Massilamany, Arunakumar Gangaplara, Heejeong Kim, Charlotte Standord, Govardhan Rathnaiah, David Steffen, Jaekwon Lee, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

In this report, we have addressed the role of copper–zinc superoxide dismutase (SOD1) deficiency in the mediation of central nervous system autoimmunity. We demonstrate that SOD1-deficient C57Bl/6 mice develop more severe autoimmune encephalomyelitis induced with myelin oligodendrocyte glycoprotein (MOG) 35–55, compared with wild type mice. This alteration in the disease phenotype was not due to aberrant expansion of MOG-specific T cells nor their ability to produce inflammatory cytokines; rather lymphocytes generated in SOD1-deficient mice were more prone to spontaneous cell death when compared with their wild type littermate controls. The data point to a role for SOD1 in the maintenance …

Rhodotorula Minuta Fungemia In A Ewe Lamb, C G. Chitko-Mckown, K A. Leymaster, M. P. Heaton, D D. Griffin, J K. Veatch, S A. Jones, M. L. Clawson

School of Veterinary and Biomedical Sciences: Faculty Publications

An 8-month-old crossbred ewe, normal upon physical examination, was humanely euthanized for tissue collection. After approximately 3 weeks in tissue culture, fungi began budding out of cells obtained from the choroid plexus. After an additional 3 weeks, budding was observed in kidney cell cultures and eventually in monocyte cultures as well. Serum from the lamb was submitted to the Veterinary Diagnostic Laboratory at Colorado State University for fungal diagnosis and was found negative for Aspergillus, Blastomyces, Coccidioidomycosis and Histoplasmosis. DNA was isolated from fungi collected from tissue culture supernatants and used in a set of pan-fungal PCR assays with DNA …

Identification Of Amino Acid Residues Important For Anti-Ifn Activity Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1, Lalit Beura, Sakthivel Subramaniam, Hiep Vu, Byungjoon Kwon, Asit K. Pattnaik, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

The non-structural protein 1 (nsp1) of porcine reproductive and respiratory syndrome virus is partly responsible for inhibition of type I interferon (IFN) response by the infected host. By performing alanine-scanning mutagenesis, we have identified amino acid residues in nsp1α and nsp1β~ (the proteolytic products of nsp1) that when substituted with alanine(s) exhibited significant relief of IFNsuppression. A mutant virus (16-SA, in which residues 16-20 of nsp1β were substituted with alanines) encoding mutant nsp1β recovered from infectious cDNA clone was shown to be attenuated for growth in vitro and induced significantly higher amount of type I IFN transcripts in infected macrophages. …

A Single Amino Acid Change Resulting In Loss Of Fluorescence Of Egfp In A Viral Fusion Protein Confers Fitness And Growth Advantage To The Recombinant Vesicular Stomatitis Virus, Phat X. Dinh, Debasis Panda, Phani B. Das, Subash C. Das, Anshuman Das, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation …

Fusion Of A Fluorescent Protein To The Pul25 Minor Capsid Protein Of Pseudorabies Virus Allows Live-Cell Capsid Imaging With Negligible Impact On Infection, Kevin P. Bohannon, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

In order to resolve the location and activity of submicroscopic viruses in living cells, viral proteins are often fused to fluorescent proteins (FPs) and visualized by microscopy. In this study, we describe the fusion of FPs to three proteins of pseudorabies virus (PRV) that allowed imaging of capsids in living cells. Included in this study are the first recombinant PRV strains expressing FP–pUL25 fusions based on a design applied to herpes simplex virus type 1 by Homa and colleagues. The properties of each reporter virus were compared in both in vitro and in vivo infection models. PRV strains expressing FP–pUL25 …