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Strand-Specific Libraries For High Throughput Rna Sequencing (Rna-Seq) Prepared Without Poly(A) Selection, Zhao Zhang, William E. Theurkauf, Zhiping Weng, Phillip D. Zamore 2017 University of Massachusetts Medical School

Strand-Specific Libraries For High Throughput Rna Sequencing (Rna-Seq) Prepared Without Poly(A) Selection, Zhao Zhang, William E. Theurkauf, Zhiping Weng, Phillip D. Zamore

Zhao Zhang

BACKGROUND: High throughput DNA sequencing technology has enabled quantification of all the RNAs in a cell or tissue, a method widely known as RNA sequencing (RNA-Seq). However, non-coding RNAs such as rRNA are highly abundant and can consume >70% of sequencing reads. A common approach is to extract only polyadenylated mRNA; however, such approaches are blind to RNAs with short or no poly(A) tails, leading to an incomplete view of the transcriptome. Another challenge of preparing RNA-Seq libraries is to preserve the strand information of the RNAs. DESIGN: Here, we describe a procedure for preparing RNA-Seq libraries from 1 ...


An Approach To Identify Mycobacteriophage Diversity Prior To Dna Sequencing, Charles Gregory 2017 Western Kentucky University

An Approach To Identify Mycobacteriophage Diversity Prior To Dna Sequencing, Charles Gregory

Honors College Capstone Experience/Thesis Projects

Over 6,869 Mycobacteriophages have been isolated and purified. Of these, 1,367 genomes have been sequenced at the DNA level and more are added each year through the SEA-PHAGES program. Sequenced mycobacteriophages are grouped into clusters based on a 50% or greater nucleotide identity. The number and breadth of these clusters represents the diversity present in the environment. Each year, as new phages are discovered by students in the SEA-PHAGES program, the question arises, “Which isolates should we sequence?” In order to sequence phages that represent the greatest possible diversity, and thus broaden under-represented clusters and identify new singletons ...


Insights Into Diterpene Cyclization From Structure Of Bifunctional Abietadiene Synthase From Abies Grandis, Ke Zhou, Yang Gao, Julie A. Hoy, Francis M. Mann, Richard B. Honzatko, Reuben J. Peters 2017 Iowa State University

Insights Into Diterpene Cyclization From Structure Of Bifunctional Abietadiene Synthase From Abies Grandis, Ke Zhou, Yang Gao, Julie A. Hoy, Francis M. Mann, Richard B. Honzatko, Reuben J. Peters

Reuben J. Peters

Abietadiene synthase from Abies grandis (AgAS) is a model system for diterpene synthase activity, catalyzing class I (ionization-initiated) and class II (protonation-initiated) cyclization reactions. Reported here is the crystal structure of AgAS at 2.3 Å resolution and molecular dynamics simulations of that structure with and without active site ligands. AgAS has three domains (α, β, and γ). The class I active site is within the C-terminal α domain, and the class II active site is between the N-terminal γ and β domains. The domain organization resembles that of monofunctional diterpene synthases and is consistent with proposed evolutionary origins of ...


Diterpene Cyclases And The Nature Of The Isoprene Fold, Rong Cao, Yonghui Zhang, Francis M. Mann, Cancan Huang, Dushyant Mukkamala, Michael P. Hudock, Matthew Mead, Sladjana Prisic, Ke Wang, Fu-Yang Lin, Ting-Kai Chang, Reuben Peters, Eric Oldfield 2017 University of Illinois at Urbana-Champaign

Diterpene Cyclases And The Nature Of The Isoprene Fold, Rong Cao, Yonghui Zhang, Francis M. Mann, Cancan Huang, Dushyant Mukkamala, Michael P. Hudock, Matthew Mead, Sladjana Prisic, Ke Wang, Fu-Yang Lin, Ting-Kai Chang, Reuben Peters, Eric Oldfield

Reuben J. Peters

The structures and mechanism of action of many terpene cyclases are known, but there are no structures of diterpene cyclases. Here, we propose structural models based on bioinformatics, sitedirected mutagenesis, domain swapping, enzyme inhibition and spectroscopy that help explain the nature of diterpene cyclase structure, function, and evolution. Bacterial diterpene cyclases contain ~20 α-helices and the same conserved “QW” and DxDD motifs as in triterpene cyclases, indicating the presence of a βγ barrel structure. Plant diterpene cyclases have a similar catalytic motif and βγ-domain structure together with a third, α-domain, forming an αβγ structure, and in H+-initiated cyclases, there ...


Microbiome Analyses Of Pacific White Shrimp (Litopenaeus Vannamei) Collected From Disparate Geographical Locations, Arun S. Seetharam, Emily Kawaler, Zhi-Qiang Du, Max F. Rothschild, Andrew J. Severin 2017 Iowa State University

Microbiome Analyses Of Pacific White Shrimp (Litopenaeus Vannamei) Collected From Disparate Geographical Locations, Arun S. Seetharam, Emily Kawaler, Zhi-Qiang Du, Max F. Rothschild, Andrew J. Severin

Max Rothschild

In this study, the tail muscle microbiota of pacific white shrimp (Litopenaeus vannamei) sourced from five countries across Central and South America and Southeast Asia were determined and compared. The genomic DNA was sequenced at around 10 × coverage for each geographical location and was assembled de novo for comparative analysis. The assembled sequences for all the lines were classified based on their similarity to the sequences in the public database. We found that there is high correlation among the microbiota of shrimp from disparate regions, as well as the presence of some DNA from bacteria known to cause food poisoning ...


Crispr/Cas9-Mediated Genome Editing Induces Exon Skipping By Alternative Splicing Or Exon Deletion, Haiwei Mou, Jordan L. Smith, Lingtao Peng, Jill Moore, Xiao-Ou Zhang, Chun-Qing Song, Ankur Sheel, Deniz M. Ozata, Yingxiang Li, Charles P. Emerson Jr., Erik J. Sontheimer, Melissa J. Moore, Zhiping Weng, Wen Xue 2017 University of Massachusetts Medical School

Crispr/Cas9-Mediated Genome Editing Induces Exon Skipping By Alternative Splicing Or Exon Deletion, Haiwei Mou, Jordan L. Smith, Lingtao Peng, Jill Moore, Xiao-Ou Zhang, Chun-Qing Song, Ankur Sheel, Deniz M. Ozata, Yingxiang Li, Charles P. Emerson Jr., Erik J. Sontheimer, Melissa J. Moore, Zhiping Weng, Wen Xue

Program in Bioinformatics and Integrative Biology Publications and Presentations

CRISPR is widely used to disrupt gene function by inducing small insertions and deletions. Here, we show that some single-guide RNAs (sgRNAs) can induce exon skipping or large genomic deletions that delete exons. For example, CRISPR-mediated editing of beta-catenin exon 3, which encodes an autoinhibitory domain, induces partial skipping of the in-frame exon and nuclear accumulation of beta-catenin. A single sgRNA can induce small insertions or deletions that partially alter splicing or unexpected larger deletions that remove exons. Exon skipping adds to the unexpected outcomes that must be accounted for, and perhaps taken advantage of, in CRISPR experiments.


Transcriptomic Analysis Of Carboxylic Acid Challenge In Escherichia Coli: Beyond Membrane Damage, Liam A. Royce, Erin E. Boggess, Yao Fu, Ping Liu, Jacqueline V. Shanks, Julie A. Dickerson, Laura R. Jarboe 2017 Iowa State University

Transcriptomic Analysis Of Carboxylic Acid Challenge In Escherichia Coli: Beyond Membrane Damage, Liam A. Royce, Erin E. Boggess, Yao Fu, Ping Liu, Jacqueline V. Shanks, Julie A. Dickerson, Laura R. Jarboe

Laura R. Jarboe

Carboxylic acids are an attractive biorenewable chemical. Enormous progress has been made in engineering microbes for production of these compounds though titers remain lower than desired. Here we used transcriptome analysis of Escherichia coli during exogenous challenge with octanoic acid (C8) at pH 7.0 to probe mechanisms of toxicity. This analysis highlights the intracellular acidification and membrane damage caused by C8 challenge. Network component analysis identified transcription factors with altered activity including GadE, the activator of the glutamate-dependent acid resistance system (AR2) and Lrp, the amino acid biosynthesis regulator. The intracellular acidification was quantified during exogenous challenge, but was ...


Membrane Stress Caused By Octanoic Acid In Saccharomyces Cerevisiae, Ping Liu, Andriy A. Chernyshova, Tarek Najdi, Yao Fu, Julie Dickerson, Suzanne Sandmeyer, Laura R. Jarboe 2017 Iowa State University

Membrane Stress Caused By Octanoic Acid In Saccharomyces Cerevisiae, Ping Liu, Andriy A. Chernyshova, Tarek Najdi, Yao Fu, Julie Dickerson, Suzanne Sandmeyer, Laura R. Jarboe

Laura R. Jarboe

In order to compete with petroleum-based fuel and chemicals, engineering a robust biocatalyst that can convert renewable feedstocks into biorenewable chemicals, such as carboxylic acids, is increasingly important. However, product toxicity is often problematic. In this study, the toxicity of the carboxylic acids hexanoic, octanoic, and decanoic acid on Saccharomyces cerevisiae was investigated, with a focus on octanoic acid. These compounds are completely inhibitory at concentrations of magnitude 1 mM, and the toxicity increases as chain length increases and as media pH decreases. Transciptome analysis, reconstruction of gene regulatory network, and network component analysis suggested decreased membrane integrity during challenge ...


Using Phylogenetically-Informed Annotation (Pia) To Search For Light-Interacting Genes In Transcriptomes From Non-Model Organisms, Daniel I. Speiser, M. Sabrina Pankey, Alexander K. Zaharoff, Barbara A. Battelle, Heather D. Bracken-Grissom, Jesse W. Breinholt, Seth M. Bybee, Thomas W. Cronin, Anders Garm, Annie R. Lindgren, Nipam H. Patel, Megan L. Porter, Meredith E. Protas, Anja S. Rivera, Jeanne M. Serb, Kirk S. Zigler, Keith A. Crandall, Todd H. Oakley 2017 Department of Ecology, Evolution, and Marine Biology, University of California Santa Barbara

Using Phylogenetically-Informed Annotation (Pia) To Search For Light-Interacting Genes In Transcriptomes From Non-Model Organisms, Daniel I. Speiser, M. Sabrina Pankey, Alexander K. Zaharoff, Barbara A. Battelle, Heather D. Bracken-Grissom, Jesse W. Breinholt, Seth M. Bybee, Thomas W. Cronin, Anders Garm, Annie R. Lindgren, Nipam H. Patel, Megan L. Porter, Meredith E. Protas, Anja S. Rivera, Jeanne M. Serb, Kirk S. Zigler, Keith A. Crandall, Todd H. Oakley

Jeanne M Serb

Background: Tools for high throughput sequencing and de novo assembly make the analysis of transcriptomes (i.e. the suite of genes expressed in a tissue) feasible for almost any organism. Yet a challenge for biologists is that it can be difficult to assign identities to gene sequences, especially from non-model organisms. Phylogenetic analyses are one useful method for assigning identities to these sequences, but such methods tend to be time-consuming because of the need to re-calculate trees for every gene of interest and each time a new data set is analyzed. In response, we employed existing tools for phylogenetic analysis ...


Transcriptomic Analysis Of Carboxylic Acid Challenge In Escherichia Coli: Beyond Membrane Damage, Liam A. Royce, Erin E. Boggess, Yao Fu, Ping Liu, Jacqueline V. Shanks, Julie A. Dickerson, Laura R. Jarboe 2017 Iowa State University

Transcriptomic Analysis Of Carboxylic Acid Challenge In Escherichia Coli: Beyond Membrane Damage, Liam A. Royce, Erin E. Boggess, Yao Fu, Ping Liu, Jacqueline V. Shanks, Julie A. Dickerson, Laura R. Jarboe

Jacqueline V. Shanks

Carboxylic acids are an attractive biorenewable chemical. Enormous progress has been made in engineering microbes for production of these compounds though titers remain lower than desired. Here we used transcriptome analysis of Escherichia coli during exogenous challenge with octanoic acid (C8) at pH 7.0 to probe mechanisms of toxicity. This analysis highlights the intracellular acidification and membrane damage caused by C8 challenge. Network component analysis identified transcription factors with altered activity including GadE, the activator of the glutamate-dependent acid resistance system (AR2) and Lrp, the amino acid biosynthesis regulator. The intracellular acidification was quantified during exogenous challenge, but was ...


Transcriptome Profiling Of Soybean (Glycine Max) Roots Challenged With Pathogenic And Non-Pathogenic Isolates Of Fusarium Oxysporum, Alessandra Lanubile, Usha K. Muppirala, Andrew J. Severin, Adriano Marocco, Gary P. Munkvold 2017 Iowa State University

Transcriptome Profiling Of Soybean (Glycine Max) Roots Challenged With Pathogenic And Non-Pathogenic Isolates Of Fusarium Oxysporum, Alessandra Lanubile, Usha K. Muppirala, Andrew J. Severin, Adriano Marocco, Gary P. Munkvold

Gary P. Munkvold

Background Fusarium oxysporum is one of the most common fungal pathogens causing soybean root rot and seedling blight in U.S.A. In a recent study, significant variation in aggressiveness was observed among isolates of F. oxysporum collected from roots in Iowa, ranging from highly pathogenic to weakly or non-pathogenic isolates. Results We used RNA-seq analysis to investigate the molecular aspects of the interactions of a partially resistant soybean genotype with non-pathogenic/pathogenic isolates of F. oxysporum at 72 and 96 h post inoculation (hpi). Markedly different gene expression profiles were observed in response to the two isolates. A peak ...


Darwin Core Archive File, Stover-Ebinger Herbarium, Eastern Illinois University 2017 Eastern Illinois University

Darwin Core Archive File, Stover-Ebinger Herbarium, Eastern Illinois University

Darwin Core Archive Download

ZIP file contains occurrences.csv, identivications.csv, and images.csv. The meta.xml document describes the content. Fields within the occurrences.csv file are defined by the Darwin Core exchange standard.


Surveillance For Sulfadoxine-Pyrimethamine Resistant Malaria Parasites In The Lake And Southern Zones, Tanzania, Using Pooling And Next-Generation Sequencing, Jeremiah M. Ngondi, Nicholas J. Hathaway, Jeffrey A. Bailey, Julie Gutman 2017 RTI International

Surveillance For Sulfadoxine-Pyrimethamine Resistant Malaria Parasites In The Lake And Southern Zones, Tanzania, Using Pooling And Next-Generation Sequencing, Jeremiah M. Ngondi, Nicholas J. Hathaway, Jeffrey A. Bailey, Julie Gutman

Program in Bioinformatics and Integrative Biology Publications and Presentations

BACKGROUND: Malaria in pregnancy (MiP) remains a major public health challenge in areas of high malaria transmission. Intermittent preventive treatment in pregnancy (IPTp) with sulfadoxine-pyrimethamine (SP) is recommended to prevent the adverse consequences of MiP. The effectiveness of SP for IPTp may be reduced in areas where the dhps581 mutation (a key marker of high level SP resistance) is found; this mutation was previously reported to be common in the Tanga Region of northern Tanzania, but there are limited data from other areas. The frequency of molecular markers of SP resistance was investigated in malaria parasites from febrile patients at ...


Statistical Analysis Of Real-Time Pcr Data, Joshua S. Yuan, Ann Reed, Feng Chen, C. Neal Stewart 2017 University of Tennessee - Knoxville

Statistical Analysis Of Real-Time Pcr Data, Joshua S. Yuan, Ann Reed, Feng Chen, C. Neal Stewart

Ann Reed

Background Even though real-time PCR has been broadly applied in biomedical sciences, data processing procedures for the analysis of quantitative real-time PCR are still lacking; specifically in the realm of appropriate statistical treatment. Confidence interval and statistical significance considerations are not explicit in many of the current data analysis approaches. Based on the standard curve method and other useful data analysis methods, we present and compare four statistical approaches and models for the analysis of real-time PCR data. Results In the first approach, a multiple regression analysis model was developed to derive ΔΔCt from estimation of interaction of gene and ...


Statistical Analysis Of Real-Time Pcr Data, Joshua S. Yuan, Ann Reed, Feng Chen, C. Neal Stewart 2017 University of Tennessee - Knoxville

Statistical Analysis Of Real-Time Pcr Data, Joshua S. Yuan, Ann Reed, Feng Chen, C. Neal Stewart

Ann Reed

Background Even though real-time PCR has been broadly applied in biomedical sciences, data processing procedures for the analysis of quantitative real-time PCR are still lacking; specifically in the realm of appropriate statistical treatment. Confidence interval and statistical significance considerations are not explicit in many of the current data analysis approaches. Based on the standard curve method and other useful data analysis methods, we present and compare four statistical approaches and models for the analysis of real-time PCR data. Results In the first approach, a multiple regression analysis model was developed to derive ΔΔCt from estimation of interaction of gene and ...


Proline Isomerization Preorganizes The Itk Sh2 Domain For Binding The Itk Sh3 Domain, Andrew J. Severin, Raji E. Joseph, Scott E. Boyken, D. Bruce Fulton, Amy H. Andreotti 2017 Iowa State University

Proline Isomerization Preorganizes The Itk Sh2 Domain For Binding The Itk Sh3 Domain, Andrew J. Severin, Raji E. Joseph, Scott E. Boyken, D. Bruce Fulton, Amy H. Andreotti

Amy Andreotti

We report here the NMR-derived structure of the binary complex formed by the interleukin-2 tyrosine kinase (Itk) Src homology 3 (SH3) and Src homology 2 (SH2) domains. The interaction is independent of both a phosphotyrosine motif and a proline-rich sequence, the classical targets of the SH2 and SH3 domains, respectively. The Itk SH3/SH2 structure reveals the molecular details of this nonclassical interaction and provides a clear picture for how the previously described prolyl cis/trans isomerization present in the Itk SH2 domain mediates SH3 binding. The higher-affinity cis SH2 conformer is preorganized to form a hydrophobic interface with the ...


Characterization Of Zic2 As An Oncoprotein In Prostate Cancer, Keira C. Davis 2017 Clark Atlanta University

Characterization Of Zic2 As An Oncoprotein In Prostate Cancer, Keira C. Davis

Electronic Theses & Dissertations Collection for Atlanta University & Clark Atlanta University

The field of prostate cancer research is in need of biological markers that predict which cancers do not need treatment, those that can be treated successfully with a localized treatment and more specific cases in which patients are likely to have an aggressive form of cancer that will require more aggressive surgical and chemotherapeutic treatments. ZIC2 is one of five members of a family of proteins that play critical roles in neural crest and mesoderm growth in normal embryonic brain development and in the adult cerebellum of vertebrates. Found throughout the animal kingdom, ZIC1-5 genes encode five distinct ZIC proteins ...


Isolation And Comparative Genomic Analysis Of Final Third Of Satis Genome, Kelly Hartigan, Nicole Curnutt, Matthew McDermut 2017 Washington University in St. Louis

Isolation And Comparative Genomic Analysis Of Final Third Of Satis Genome, Kelly Hartigan, Nicole Curnutt, Matthew Mcdermut

Undergraduate Research Symposium Posters

A highly novel Streptomyces phage, Satis, was isolated from a direct environmental sample collected from outside Danforth House on the Washington University campus. Satis infects bacterial species Streptomyces lividans producing pinpoint, cloudy plaques less than 1mm in diameter. Electron microscope data shows rare atypical physical features. Rather than the common octahedral capsid shape, Satis has a prolate head with visible cross-linked hexagonal protein structure and average measurements of 285 nm by 47 nm with a long, flexible tail measuring 268 nm. Upon sequencing, it was found that Satis contains the longest phage genome discovered to date through the SEA-PHAGE program ...


Evolutionary Genetic Aspects Of Host Association In Generalist Ectoparasites, Benoit Talbot 2017 The University of Western Ontario

Evolutionary Genetic Aspects Of Host Association In Generalist Ectoparasites, Benoit Talbot

Electronic Thesis and Dissertation Repository

Despite the use of the host for dispersal by most parasite species, the extremely loose relationship typical between highly mobile hosts and generalist ectoparasites may lead to very different gene flow patterns between the two, leading in turn to different spatial genetic structure, and potentially different demographic history. I examined how similar gene flow patterns are between Cimex adjunctus, a generalist ectoparasite of bats present throughout North America, and two of its key bat hosts. I first analyzed the continent-scale genetic structure and demographic history of C. adjunctus and compared it to that of two of its hosts, the little ...


Can Cone Signals In The Wild Be Predicted From The Past?, David H. Foster, Iván Marín-Franch 2017 University of Manchester, UK

Can Cone Signals In The Wild Be Predicted From The Past?, David H. Foster, Iván Marín-Franch

MODVIS Workshop

In the natural world, the past is usually a good guide to the future. If light from the sun and sky is blue earlier in the day and yellow now, then it is likely to be more yellow later, as the sun's elevation decreases. But is the light reflected from a scene into the eye as predictable as the light incident upon the scene, especially when lighting changes are not just spectral but include changes in local shadows and mutual reflections? The aim of this work was to test the predictability of cone photoreceptor signals in the wild over ...


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