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2004

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Full-Text Articles in Life Sciences

Chromosomal Duplications And Cointegrates Generated By The Bacteriophage Lamdba Red System In Escherichia Coli K-12, Anthony R. Poteete, Anita C. Fenton, Ashwini Nadkarni Dec 2004

Chromosomal Duplications And Cointegrates Generated By The Bacteriophage Lamdba Red System In Escherichia Coli K-12, Anthony R. Poteete, Anita C. Fenton, Ashwini Nadkarni

Open Access Articles

BACKGROUND: An Escherichia coli strain in which RecBCD has been genetically replaced by the bacteriophage lambda Red system engages in efficient recombination between its chromosome and linear double-stranded DNA species sharing sequences with the chromosome. Previous studies of this experimental system have focused on a gene replacement-type event, in which a 3.5 kbp dsDNA consisting of the cat gene and flanking lac operon sequences recombines with the E. coli chromosome to generate a chloramphenicol-resistant Lac- recombinant. The dsDNA was delivered into the cell as part of the chromosome of a non-replicating lambda vector, from which it was released by ...


The Histone Fold Domain Of Cse4 Is Sufficient For Cen Targeting And Propagation Of Active Centromeres In Budding Yeast, Lisa Morey, Kelly Barnes, Yinhuai Chen, Molly Fitzgerald-Hayes, Richard E. Baker Dec 2004

The Histone Fold Domain Of Cse4 Is Sufficient For Cen Targeting And Propagation Of Active Centromeres In Budding Yeast, Lisa Morey, Kelly Barnes, Yinhuai Chen, Molly Fitzgerald-Hayes, Richard E. Baker

Open Access Articles

Centromere-specific H3-like proteins (CenH3s) are conserved across the eukaryotic kingdom and are required for packaging centromere DNA into a specialized chromatin structure required for kinetochore assembly. Cse4 is the CenH3 protein of the budding yeast Saccharomyces cerevisiae. Like all CenH3 proteins, Cse4 consists of a conserved histone fold domain (HFD) and a divergent N terminus (NT). The Cse4 NT contains an essential domain designated END (for essential N-terminal domain); deletion of END is lethal. To investigate the role of the Cse4 NT in centromere targeting, a series of deletion alleles (cse4DeltaNT) were analyzed. No part of the Cse4 NT was ...


Allele Specific Synthetic Lethality Between Pric And Dnaats Alleles At The Permissive Temperature Of 30°C In E. Coli K-12, Steven Sandler, Tania Hinds Dec 2004

Allele Specific Synthetic Lethality Between Pric And Dnaats Alleles At The Permissive Temperature Of 30°C In E. Coli K-12, Steven Sandler, Tania Hinds

Steven Sandler

Background DnaA is an essential protein in the regulation and initiation of DNA replication in many bacteria. It forms a protein-DNA complex at oriC to which DnaC loads DnaB. DNA replication forks initiated at oriC by DnaA can collapse on route to the terminus for a variety of reasons. PriA, PriB, PriC, DnaT, Rep and DnaC form multiple pathways to restart repaired replication forks. DnaC809 and dnaC809,820 are suppressors of priA2::kan mutant phenotypes. The former requires PriC and Rep while the latter is independent of them. RnhA339::cat mutations allow DnaA-independent initiation of DNA replication. Results It is ...


Translational Regulation Of Gene Expression, Stephanie Kervestin, Nadia Amrani Dec 2004

Translational Regulation Of Gene Expression, Stephanie Kervestin, Nadia Amrani

Open Access Articles

A report on the Cold Spring Harbor Laboratory meeting 'Translational Control', Cold Spring Harbor, USA, 7-12 September 2004.


Identification Of Proteins Involved In Chloroplast Dna Replication, Matthew G. Lassen Dec 2004

Identification Of Proteins Involved In Chloroplast Dna Replication, Matthew G. Lassen

Theses and Dissertations

Chapter 1

Chloroplast nucleoids (ct-nucleoids) are DNA/protein complexes involved in compacting the chloroplast genome, and may play a role in regulating DNA replication. Ct-nucleoids were isolated from young soybean plants and separated by 2-D gel electrophoresis. Gel spots were excised and analyzed by MALDI-ToF mass spectrometry, resulting in several protein identifications. The proteins identified all have functions unrelated to DNA replication. While some of these proteins may be due to contamination, it is possible that some of these proteins are dual-functional, playing direct roles in the regulation of DNA replication.

Chapter 2

A 28 kDa soybean protein was isolated ...


Dna Microarray Analysis Of Nitrogen Fixation And Fe(Iii) Reduction In Geobacter Sulfurreducens, Jennifer Webster, Barbara A. Methé, Kelly P. Nevin, Jessica E. Butler, Derek Lovley Dec 2004

Dna Microarray Analysis Of Nitrogen Fixation And Fe(Iii) Reduction In Geobacter Sulfurreducens, Jennifer Webster, Barbara A. Methé, Kelly P. Nevin, Jessica E. Butler, Derek Lovley

Derek Lovley

A DNA microarray representing the genome of Geobacter sulfurreducens was constructed for use in global gene expression profiling of cells under steady-state conditions with acetate as the electron donor and Fe(III) or fumarate as the electron acceptor. Reproducible differences in transcript levels were also observed in comparisons between cells grown with ammonia and those fixing atmospheric nitrogen. There was a high correlation between changes in transcript levels determined with microarray analyses and an evaluation of a subset of the genome with quantitative PCR. As expected, cells required to fix nitrogen had higher levels of transcripts of genes associated with ...


In Situ Expression Of Nigd In Geobacteraceae In Subsurface Sediments, Dawn E. Holmes, Kelly P. Nevin, Derek Lovley Dec 2004

In Situ Expression Of Nigd In Geobacteraceae In Subsurface Sediments, Dawn E. Holmes, Kelly P. Nevin, Derek Lovley

Kelly Nevin

In order to determine whether the metabolic state of Geobacteraceae involved in bioremediation of subsurface sediments might be inferred from levels of mRNA for key genes, in situ expression of nifD, a highly conserved gene involved in nitrogen fixation, was investigated. When Geobacter sulfurreducens was grown without a source of fixed nitrogen in chemostats with acetate provided as the limiting electron donor and Fe(III) as the electron acceptor, levels of nifD transcripts were 4 to 5 orders of magnitude higher than in chemostat cultures provided with ammonium. In contrast, the number of transcripts of recA and the 16S rRNA ...


Infectious Molecular Clone Of A Recently Transmitted Pediatric Human Immunodeficiency Virus Clade C Isolate From Africa: Evidence Of Intraclade Recombination, Ricky D. Grisson, Agnes-Laurence Chenine, Lan-Yu Yeh, Jun He, Charles Wood, Ganapati J. Bhat, Weidong Zu, Chipepo Kankasa, Ruth M. Ruprecht Dec 2004

Infectious Molecular Clone Of A Recently Transmitted Pediatric Human Immunodeficiency Virus Clade C Isolate From Africa: Evidence Of Intraclade Recombination, Ricky D. Grisson, Agnes-Laurence Chenine, Lan-Yu Yeh, Jun He, Charles Wood, Ganapati J. Bhat, Weidong Zu, Chipepo Kankasa, Ruth M. Ruprecht

Virology Papers

Although human immunodeficiency virus type 1 (HIV-1) clade C continues to dominate the pandemic, only two infectious clade C proviral DNA clones have been described (N. Mochizuki, N. Otsuka, K. Matsuo, T. Shiino, A. Kojima, T. Kurata, K. Sakai, N. Yamamoto, S. Isomura, T. N. Dhole, Y. Takebe, M. Matsuda, and M. Tatsumi, AIDS Res. Hum. Retrovir. 15:1321–1324, 1999; T. Ndung’u, B. Renjifo, and M. Essex, J. Virol. 75:4964–4972, 2001). We have generated an infectious molecular clone of a pediatric clade C strain, HIV1084i, which was isolated from a Zambian infant infected either intrapartum or ...


Interferon Regulatory Factor 7 Is Associated With Epstein-Barr Virus-Transformed Central Nervous System Lymphoma And Has Oncogenic Properties, Luwen Zhang, Jun Zhang, Que Lambert, Channing J. Der, Luis Del Valle, Judith Miklossy, Kamel Kahlili, You Zhou, Joseph S. Pagano Dec 2004

Interferon Regulatory Factor 7 Is Associated With Epstein-Barr Virus-Transformed Central Nervous System Lymphoma And Has Oncogenic Properties, Luwen Zhang, Jun Zhang, Que Lambert, Channing J. Der, Luis Del Valle, Judith Miklossy, Kamel Kahlili, You Zhou, Joseph S. Pagano

Virology Papers

EBV transforms primary B cells, and the major EBV oncoprotein, latent membrane protein 1 (LMP-1), is required for the process. LMP-1 both induces the expression of IRF-7 and activates the IRF-7 protein by phosphorylation and nuclear translocation. Here we report that the expression of IRF-7 is increased in EBV-immortalized B lymphocytes compared with that in primary B cells. IRF-7 was phosphorylated and predominantly localized in the nucleus in the immortalized cells. The expression of IRF-7 was detected in 19 of 27 specimens of primary lymphomas of the human central nervous system by immunohistochemical analysis. The association between LMP-1 and IRF-7 ...


Association Of Dc-Sign Promoter Polymorphism With Increased Risk For Parenteral, But Not Mucosal, Acquisition Of Human Immunodeficiency Virus Type 1 Infection, Maureen P. Martin, Michael M. Lederman, Holli B. Hutcheson, James J. Goedert, George W. Nelson, Yvette Van Kooyk, Roger Detels, Susan Buchbinder, Keith Hoots, David Vlahov, Stephen J. O'Brien, Mary Carrington Dec 2004

Association Of Dc-Sign Promoter Polymorphism With Increased Risk For Parenteral, But Not Mucosal, Acquisition Of Human Immunodeficiency Virus Type 1 Infection, Maureen P. Martin, Michael M. Lederman, Holli B. Hutcheson, James J. Goedert, George W. Nelson, Yvette Van Kooyk, Roger Detels, Susan Buchbinder, Keith Hoots, David Vlahov, Stephen J. O'Brien, Mary Carrington

Biology Faculty Articles

There is considerable debate about the fundamental mechanisms that underlie and restrict acquisition of human immunodeficiency virus type 1 (HIV-1) infection. In light of recent studies demonstrating the ability of C type lectins to facilitate infection with HIV-1, we explored the potential relationship between polymorphisms in the DC-SIGN promoter and risk for acquisition of HIV-1 according to route of infection. Using samples obtained from 1,611 European-American participants at risk for parenteral (n = 713) or mucosal (n = 898) infection, we identified single-nucleotide polymorphisms in the DC-SIGN promoter using single-strand conformation polymorphism. Individuals at risk for parenterally acquired infection who had ...


Using A Bioluminescent Bacterial Bioreporter To Assess Iron Bioavailability In The Oceans, Cecile Elise Mioni Dec 2004

Using A Bioluminescent Bacterial Bioreporter To Assess Iron Bioavailability In The Oceans, Cecile Elise Mioni

Doctoral Dissertations

Recent improvements in modern analytical methods have considerably increased our understanding of Fe biogeochemistry in the Ocean. Compiled data have shown that Fe concentrations are low in most open ocean surface waters and that the bioavailability of this Fe is influenced by organic complexation. Of presumed biotic origin, the importance of this organic complexation to Fe availability remains to be elucidated. Unfortunately, current analytical tools do not allow for a linkage to be established between Fe speciation to Fe bioavailability.

To supplement chemical analyses, we have developed a bioanalytical tool: a heterotrophic bacterial bioluminescent reporter system that responds quantitatively to ...


Electrotransformation Of Clostridium Thermocellum, Michael V. Tyurin, Sunil G. Desai, Lee R. Lynd Nov 2004

Electrotransformation Of Clostridium Thermocellum, Michael V. Tyurin, Sunil G. Desai, Lee R. Lynd

Open Dartmouth: Faculty Open Access Scholarship

Electrotransformation of several strains of Clostridium thermocellum was achieved using plasmid pIKm1 with selection based on resistance to erythromycin and lincomycin. A custom-built pulse generator was used to apply a square 10-ms pulse to an electrotransformation cuvette consisting of a modified centrifuge tube. Transformation was verified by recovery of the shuttle plasmid pIKm1 from presumptive transformants of C. thermocellum with subsequent PCR specific to the mls gene on the plasmid, as well as by retransformation of Escherichia coli. Optimization carried out with strain DSM 1313 increased transformation efficiencies from <1 to (2.2 􏰍 0.5) 􏰎 105 transformants per 􏰃g of plasmid DNA. Factors conducive to achieving high transformation efficiencies included optimized periods of incubation both before and after electric pulse application, chilling during cell collection and washing, subculture in the presence of isoniacin prior to electric pulse application, a custom-built cuvette embedded in an ice block during pulse application, use of a high (25-kV/cm) field strength, and induction of the mls gene before plating the cells on selective medium. The protocol and preferred conditions developed for strain DSM 1313 resulted in transformation efficiencies of (5.0 􏰍 1.8) 􏰎 104 transformants per 􏰃g of plasmid DNA for strain ATCC 27405 and 􏰁1 􏰎 103 transformants per 􏰃g of plasmid DNA for strains DSM 4150 and 7072. Cell viability under optimal conditions was 􏰁50% of that of controls not exposed to an electrical pulse. Dam methylation had a beneficial but modest (7-fold for strain ATCC 27405; 40-fold for strain DSM 1313) effect on transformation efficiency. The effect of isoniacin was also strain specific. The results reported here provide for the first time a gene transfer method functional in C. thermocellum that is suitable for molecular manipulations involving either the introduction of genes associated with foreign gene products or knockout of native genes.


Evidence For Involvement Of An Electron Shuttle In Electricity Generation By Geothrix Fermentans, Daiel R. Bond, Derek Lovley Nov 2004

Evidence For Involvement Of An Electron Shuttle In Electricity Generation By Geothrix Fermentans, Daiel R. Bond, Derek Lovley

Derek Lovley

In experiments performed using graphite electrodes poised by a potentiostat (+200 mV versus Ag/AgCl) or in a microbial fuel cell (with oxygen as the electron acceptor), the Fe(III)-reducing organism Geothrix fermentans conserved energy to support growth by coupling the complete oxidation of acetate to reduction of a graphite electrode. Other organic compounds, such as lactate, malate, propionate, and succinate as well as components of peptone and yeast extract, were utilized for electricity production. However, electrical characteristics and the results of shuttling assays indicated that unlike previously described electrode-reducing microorganisms, G. fermentans produced a compound that promoted electrode ...


Dinitrogen Fixation And Release Of Ammonium And Dissolved Organic Nitrogen By Trichodesmium Ims101, Mr Mulholland, Da Bronk, Dg Capone Nov 2004

Dinitrogen Fixation And Release Of Ammonium And Dissolved Organic Nitrogen By Trichodesmium Ims101, Mr Mulholland, Da Bronk, Dg Capone

VIMS Articles

Two methods used to measure dinitrogen (N-2) fixation (acetylene reduction and N-15(2) uptake) often result in different N-2 fixation rates. Part of the discrepancy may arise from the observation that Trichodesmium can release a fraction of their recently fixed N-2 as dissolved organic nitrogen (DON) and/or ammonium (NH4+). To resolve outstanding issues regarding N-2 fixation and the production of dissolved combined nitrogen (N) by Trichodesmium, we conducted a comprehensive analysis of N-2 fixation and the production of DON and NH4+ in cultures of Trichodesmium IMS101. We performed N-15(2) uptake experiments in parallel with acetylene (C2H2) reduction assays ...


Real-Time Pcr For Detection And Quantification Of The Protistan Parasite Perkinsus Marinus In Environmental Waters, C Audemard, Ks Reece, Em Burreson Nov 2004

Real-Time Pcr For Detection And Quantification Of The Protistan Parasite Perkinsus Marinus In Environmental Waters, C Audemard, Ks Reece, Em Burreson

VIMS Articles

The protistan parasite Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica along the east coast of the United States. Very few data have been collected, however, on the abundance of the parasite in environmental waters, limiting our understanding of P. marinus transmission dynamics. Real-time PCR assays with SybrGreen I as a label for detection were developed in this study for quantification of P. marinus in environmental waters with P. marinus species-specific primers and of Perkinsus spp. with Perkinsus genus-specific primers. Detection of DNA concentrations as low as the equivalent of 3.3 x 10(-2) cell per ...


Transport Of The Harmful Bloom Alga Aureococcus Anophagefferens By Oceangoing Ships And Coastal Boats, Martina A. Doblin, Linda C. Popels, Kathryn J. Coyne, David A. Hutchins, S. Craig Cary, Fred C. Dobbs Nov 2004

Transport Of The Harmful Bloom Alga Aureococcus Anophagefferens By Oceangoing Ships And Coastal Boats, Martina A. Doblin, Linda C. Popels, Kathryn J. Coyne, David A. Hutchins, S. Craig Cary, Fred C. Dobbs

OEAS Faculty Publications

It is well established that cyst-forming phytoplankton species are transported in ships' ballast tanks. However, there is increasing evidence that other phytoplankton species which do not encyst are also capable of surviving ballast transit. These species have alternative modes of nutrition (hetero- or mixotrophy) and/or are able to survive long-term darkness. In our studies of no-ballast-on-board vessels arriving in the Great Lakes, we tested for the presence of the harmful algal bloom species Aureococcus anophagefferens (brown tide) in residual (i.e., unpumpable) ballast water using methods based on the PCR. During 2001, the brown tide organism was detected in ...


Intact Gram-Negative Helicobacter Pylori, Helicobacter Felis, And Helicobacter Hepaticus Bacteria Activate Innate Immunity Via Toll-Like Receptor 2 But Not Toll-Like Receptor 4, Leisa Mandell, Anthony P. Moran, Andrew Cocchiarella, Jeanmarie Houghton, Nancy Taylor, James G. Fox, Timothy C. Wang, Evelyn A. Kurt-Jones Nov 2004

Intact Gram-Negative Helicobacter Pylori, Helicobacter Felis, And Helicobacter Hepaticus Bacteria Activate Innate Immunity Via Toll-Like Receptor 2 But Not Toll-Like Receptor 4, Leisa Mandell, Anthony P. Moran, Andrew Cocchiarella, Jeanmarie Houghton, Nancy Taylor, James G. Fox, Timothy C. Wang, Evelyn A. Kurt-Jones

Open Access Articles

Molecular and genetic studies have demonstrated that members of the Toll-like receptor (TLR) family are critical innate immune receptors. TLRs are recognition receptors for a diverse group of microbial ligands including bacteria, fungi, and viruses. This study demonstrates that distinct TLRs are responsible for the recognition of Helicobacter lipopolysaccharide (LPS) versus intact Helicobacter bacteria. We show that the cytokine-inducing activity of Helicobacter LPS was mediated by TLR4; i.e., TLR4-deficient macrophages were unresponsive to Helicobacter pylori LPS. Surprisingly, the cytokine response to whole Helicobacter bacteria (H. pylori, H. hepaticus, and H. felis) was mediated not by TLR4 but rather by ...


The Latent Membrane Protein 1 Of Epstein-Barr Virus Establishes An Antiviral State Via Induction Of Interferon-Stimulated Genes, Jun Zhang, Subash C. Das, Catherine Kotalik, Asit K. Pattnaik, Luwen Zhang Oct 2004

The Latent Membrane Protein 1 Of Epstein-Barr Virus Establishes An Antiviral State Via Induction Of Interferon-Stimulated Genes, Jun Zhang, Subash C. Das, Catherine Kotalik, Asit K. Pattnaik, Luwen Zhang

Virology Papers

Epstein-Barr virus (EBV) infection is associated with several human cancers. Latent membrane protein 1 (LMP-1) is one of the key viral proteins required for transformation of primary B cells in vitro and establishment of EBV latency. In this report, we show that LMP-1 is able to induce the expression of several interferon (IFN)-stimulated genes (ISGs) with antiviral properties such as 2’-5’ oligoadenylate synthetase (OAS), stimulated trans-acting factor of 50 kDa (STAF- 50), and ISG-15. LMP-1 inhibits vesicular stomatitis virus (VSV) replication at low multiplicity of infection (0.1 pfu/cell). The antiviral effect of LMP-1 is associated with ...


Gel Shift Analysis Of The Empa Promoter Region In Vibrio Anguillarum, Steven M. Denkin, Pedja Sekaric, David R. Nelson Oct 2004

Gel Shift Analysis Of The Empa Promoter Region In Vibrio Anguillarum, Steven M. Denkin, Pedja Sekaric, David R. Nelson

Open Access Articles

BACKGROUND: The induction of metalloprotease encoded by empA in Vibrio anguillarum occurs at high cell density in salmon intestinal mucus. Previously we have shown that there are significant differences in empA expression in two strains of V. anguillarum, M93Sm and NB10. It is hypothesized that differences in empA regulation are due to differences in binding of regulatory elements.

RESULTS: Two strains of V. anguillarum, M93Sm and NB10, were examined and compared for the presence of DNA regulatory proteins that bind to and control the empA promoter region. Gel mobility shift assays, using a digoxigenin (DIG)-labeled oligomer containing a lux ...


Judging A Virus By Its Cover, Eva Szomolanyi-Tsuda, Raymond M. Welsh Oct 2004

Judging A Virus By Its Cover, Eva Szomolanyi-Tsuda, Raymond M. Welsh

Open Access Articles

The production of protective neutralizing antibodies occurs quickly in some viral infections but very slowly in others. In a new study, surface glycoproteins (the targets of neutralization) of 2 different viruses were genetically switched. Analysis of the neutralizing antibody response to each of the 2 parent and recombinant viruses in infected mice revealed that the speed of neutralizing antibody induction was intrinsically dependent on the surface glycoprotein and not the rest of the virus.


A Morph-Specific Daily Cycle In The Rate Of Jh Biosynthesis Underlies A Morph-Specific Daily Cycle In The Hemolymph Jh Titer In A Wing-Polymorphic Cricket, Zhangwu Zhao, Anthony J. Zera Oct 2004

A Morph-Specific Daily Cycle In The Rate Of Jh Biosynthesis Underlies A Morph-Specific Daily Cycle In The Hemolymph Jh Titer In A Wing-Polymorphic Cricket, Zhangwu Zhao, Anthony J. Zera

Anthony Zera Publications

A previous study documented a high amplitude, morph-specific daily cycle in the hemolymph JH titer in the wing-polymorphic cricket, Gryllus firmus. The JH titer rose and fell 10–20 fold in the flight-capable [LW(f), long-winged] morph during the late-photophase- early scotophase, while it was relatively constant during that time in the flightless (SW, short-winged) morph. In the present study we documented a dramatic morph-specific daily cycle in the in vitro rate of juvenile hormone (JH) biosynthesis that was tightly correlated with the hemolymph JH titer on days 5–7 of adulthood. Biosynthetic rates rose and fell 1–2 fold ...


Cd8+ T Cells Responding To Influenza Infection Reach And Persist At Higher Numbers Than Cd4+ T Cells Independently Of Precursor Frequency, Timothy J. Powell, Deborah M. Brown, Joseph A. Hollenbaugh, Tina Charbonneau, Roslyn A. Kemp, Susan L. Swain, Richard Dutton Oct 2004

Cd8+ T Cells Responding To Influenza Infection Reach And Persist At Higher Numbers Than Cd4+ T Cells Independently Of Precursor Frequency, Timothy J. Powell, Deborah M. Brown, Joseph A. Hollenbaugh, Tina Charbonneau, Roslyn A. Kemp, Susan L. Swain, Richard Dutton

Virology Papers

The activation, localization, phenotypic changes, and function of CFSE-labeled naive influenza-specific CD8+ and CD4+ T cells following influenza infection were examined. Response of adoptively transferred CD8+ T cells was seen earliest in draining lymph node. Highly activated cells were found later in the lung, airways, and spleen, were cytolytic, and expressed IFN-γ upon restimulation. Similar amounts of division at early time points, but higher numbers of CD8+ T cells, were detected at 9 and 30 days postinfection after cotransfer of CD4+ and CD8+ T cells followed by infection. Transfer of much smaller numbers of CD4+ and CD8+ T cells led ...


Detection And Quantitation Of Bovine Respiratory Syncytial Virus Using Real-Time Quantitative Rt-Pcr And Quantitative Competitive Rt-Pcr Assays, Jenna Achenbaum, Christina L. Topliff, Ventzislav B. Vassilev, Ruben O. Donis, Kent M. Eskridge, Clayton L. Kelling Oct 2004

Detection And Quantitation Of Bovine Respiratory Syncytial Virus Using Real-Time Quantitative Rt-Pcr And Quantitative Competitive Rt-Pcr Assays, Jenna Achenbaum, Christina L. Topliff, Ventzislav B. Vassilev, Ruben O. Donis, Kent M. Eskridge, Clayton L. Kelling

Virology Papers

A single tube, fluorogenic probe-based, real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay was developed for detection and quantitation of bovine respiratory syncytial virus (BRSV) using BioRad’s iCycler iQ™. Real-time Q-RT-PCR was compared with quantitative competitive RT-PCR (QC-RT-PCR) and viral titers. Viral mRNA levels were measured in BRSV-infected bovine turbinate cell lysate harvested at eight time points (1.5, 6, 12, 24, 36, 48, 60, 72 h) post-infection. A homologous BRSV cRNA standard was used for quantitation of the mRNA by plotting a standard curve of cycle threshold (Ct) values versus standard 10-fold dilutions of cRNA of known concentrations ...


Apobec3g Genetic Variants And Their Influence On The Progression To Aids, Ping An, Gabriela Bleiber, Priya Duggal, George Nelson, Margaret May, Bastien Mangeat, Irene Alobwede, Didier Trono, David Vlahov, Sharyne Donfield, James J. Goedert, John Phair, Susan Buchbinder, Stephen J. O'Brien, Amalio Telenti, Cheryl Winkler Oct 2004

Apobec3g Genetic Variants And Their Influence On The Progression To Aids, Ping An, Gabriela Bleiber, Priya Duggal, George Nelson, Margaret May, Bastien Mangeat, Irene Alobwede, Didier Trono, David Vlahov, Sharyne Donfield, James J. Goedert, John Phair, Susan Buchbinder, Stephen J. O'Brien, Amalio Telenti, Cheryl Winkler

Biology Faculty Articles

The cytosine deaminase APOBEC3G, in the absence of the human immunodeficiency virus type 1 (HIV-1) accessory gene HIV-1 viral infectivity factor (vif), inhibits viral replication by introducing G→A hypermutation in the newly synthesized HIV-1 DNA negative strand. We tested the hypothesis that genetic variants of APOBEC3G may modify HIV-1 transmission and disease progression. Single nucleotide polymorphisms were identified in the promoter region (three), introns (two), and exons (two). Genotypes were determined for 3,073 study participants enrolled in six HIV-AIDS prospective cohorts. One codon-changing variant, H186R in exon 4, was polymorphic in African Americans (AA) (f < 37%) and rare in European Americans (f < 3%) or Europeans (f = 5 ...


T-Cell Responses To The M3 Immune Evasion Protein Of Murid Gammaherpesvirus 68 Are Partially Protective And Induced With Lytic Antigen Kinetics, Joshua J. Obar, Douglas C. Donovan, Sarah G. Crist, Ondine Silvia, James P. Stewart, Edward J. Usherwood Oct 2004

T-Cell Responses To The M3 Immune Evasion Protein Of Murid Gammaherpesvirus 68 Are Partially Protective And Induced With Lytic Antigen Kinetics, Joshua J. Obar, Douglas C. Donovan, Sarah G. Crist, Ondine Silvia, James P. Stewart, Edward J. Usherwood

Open Dartmouth: Faculty Open Access Scholarship

DNA vaccination with the M3 gene, encoding an immune evasion molecule expressed during both the acute lytic and persistent phases of murid gammaherpesvirus 68 infection, yielded a significantly lower titer of virus in the lung than controls. The protection seen was dependent on T cells, and we mapped an epitope recognized by CD8 T cells. The immune response to this epitope follows the same kinetics as lytic cycle antigens, despite the fact that this gene is expressed in both lytic and persistent stages of infection. This has important implications for our understanding of T-cell responses to putative latency-associated gammaherpesvirus proteins ...


Pleomorphic Liposarcoma In A Ten Year Old Child, Z Ahmed, H U. Shah, N Yaqoob, S Muzaffar Oct 2004

Pleomorphic Liposarcoma In A Ten Year Old Child, Z Ahmed, H U. Shah, N Yaqoob, S Muzaffar

Department of Pathology and Laboratory Medicine

No abstract provided.


Comparisons Of The M1 Genome Segments And Encoded Mu2 Proteins Of Different Reovirus Isolates, Peng Yin, Natalie D. Keirstead, Teresa J. Broering, Michelle M. Arnold, John S. L. Parker, Max L. Nibert, Kevin M. Coombs Sep 2004

Comparisons Of The M1 Genome Segments And Encoded Mu2 Proteins Of Different Reovirus Isolates, Peng Yin, Natalie D. Keirstead, Teresa J. Broering, Michelle M. Arnold, John S. L. Parker, Max L. Nibert, Kevin M. Coombs

Open Access Articles

BACKGROUND: The reovirus M1 genome segment encodes the mu2 protein, a structurally minor component of the viral core, which has been identified as a transcriptase cofactor, nucleoside and RNA triphosphatase, and microtubule-binding protein. The mu2 protein is the most poorly understood of the reovirus structural proteins. Genome segment sequences have been reported for 9 of the 10 genome segments for the 3 prototypic reoviruses type 1 Lang (T1L), type 2 Jones (T2J), and type 3 Dearing (T3D), but the M1 genome segment sequences for only T1L and T3D have been previously reported. For this study, we determined the M1 nucleotide ...


Preserved Antigenicity Of Hiv-1 P24 Produced And Purified In High Yields From Plants Inoculated With A Tobacco Mosaic Virus (Tmv)-Derived Vector, D. M. Pérez-Filgueira, B. P. Brayfield, S. Phiri, M. V. Borca, Charles Wood, Thomas Jack Morris Sep 2004

Preserved Antigenicity Of Hiv-1 P24 Produced And Purified In High Yields From Plants Inoculated With A Tobacco Mosaic Virus (Tmv)-Derived Vector, D. M. Pérez-Filgueira, B. P. Brayfield, S. Phiri, M. V. Borca, Charles Wood, Thomas Jack Morris

Papers in Virology

Production of structural proteins from foot-and-mouth disease virus (FMDV) and bovine herpes virus (BHV-1) in Nicotiana benthamiana through the use of a tobacco mosaic virus-based vector (TMV-30B) has been reported previously. The development of the TMV-30B-HISc vector, a new version that adds a C-terminal histidine (His) sequence to the foreign protein expressed is described. Coding sequences from the FMDV VPl protein and the core protein, p24, from a clade C HIV-1 isolate from Zambia were cloned into the new vector and infective RNAs were generated for each construct to inoculate N. benthamiana plants. His-tagged proteins were purified from inoculated leaves ...


In Vitro Expression Of Full-Length And Truncated Bovine Respiratory Syncytial Virus G Proteins And Their Antibody Responses In Balb/C Mice, Ryan P. Brady, Christina L. Topliff, Clayton L. Kelling Sep 2004

In Vitro Expression Of Full-Length And Truncated Bovine Respiratory Syncytial Virus G Proteins And Their Antibody Responses In Balb/C Mice, Ryan P. Brady, Christina L. Topliff, Clayton L. Kelling

Virology Papers

Bovine respiratory syncytial virus (BRSV) is a primary cause of lower respiratory tract disease in calves. Protection is incomplete following vaccination or natural infection, as re-infections are common. The objectives of this study were to create plasmid DNA constructs encoding the full-length, secreted, or conserved region of the BRSV G glycoprotein, and to compare and evaluate their expression in cell culture and potential to induce antibody responses in BALB/c mice. Transfection of COS-7 cells with plasmid DNA resulted in expression of the BRSV G region from each of the plasmid DNA constructs. Following inoculation of BALB/c mice with ...


Effect Of A Juvenile Hormone Analogue On Lipid Metabolism In A Wing-Polymorphic Cricket: Implications For The Endocrine-Biochemical Bases Of Life-History Trade-Offs, Anthony J. Zera, Zhangwu Zhao Sep 2004

Effect Of A Juvenile Hormone Analogue On Lipid Metabolism In A Wing-Polymorphic Cricket: Implications For The Endocrine-Biochemical Bases Of Life-History Trade-Offs, Anthony J. Zera, Zhangwu Zhao

Anthony Zera Publications

The wing-polymorphic cricket, Gryllus firmus, has a flight-capable morph (LW[f]: long winged with functional flight muscles) and a flightless morph (SW: short winged with reduced nonfunctional flight muscles) that differ genetically in many aspects of lipid metabolism. To determine whether these differences result from genetically based alterations in endocrine regulation, the juvenile hormone mimic, methoprene, was applied to the LW(f) morph. This hormone manipulation converted the LW(f) morph into a SW phenocopy with respect to all aspects of lipid metabolism studied; that is, methoprene application decreased in vivo biosynthesis of total lipid and triglyceride, increased absolute and ...