Genetics and Genomics Commons^™

Variation At Tri- And Tetranucleotide Repeat Microsatellite Loci In The Fruit Bat Genus Cynopterus (Chiroptera: Pteropodidae), Jay F. Storz

Jay F. Storz Publications

There is considerable uncertainty surrounding the taxonomic relationship between Cynopterus sphinx and C. brachyotis, and the status of the many named forms within C. sphinx (Storz & Kunz 1999). Polymorphic microsatellite markers for cynopterine fruit bats would greatly aid efforts to elucidate species boundaries and genetic correlates of morphological variation within species. To assess levels of variation in C. sphinx and C. brachyotis, microsatellite genotypes were obtained for a total of 731 bats (620 C. sphinx and 111 C. brachyotis).

Hybridization Of Dna By Sequential Immobilization Of Oligonucleotides At The Air-Water Interface, Murali Sastry, Vidya Ramakrishnan, Mrunalini Pattarkine, Anand Gole, K. N. Ganesh

Faculty Works

The hybridization of DNA by sequential electrostatic and hydrogen-bonding immobilization of single-stranded complementary oligonucleotides at the air-water interface with cationic Langmuir monolayers is demonstrated. The complexation of the single-stranded DNA molecules with octadecylamine (ODA) Langmuir monolayers was followed in time by monitoring the pressure-area isotherms. A large (and slow) expansion of the ODA monolayer was observed during each stage of complexation in the following sequence: primary single-stranded DNA followed by complementary single-stranded DNA followed by the intercalator, ethidium bromide. Langmuir-Blodgett (LB) films of the ODA-DNA complex were formed on different substrates and characterized using quartz-crystal microgravimetry (QCM), Fourier transform infrared …

Actin Gene Family Evolution And The Phylogeny Of Coleoid Cephalopods (Mollusca: Cephalopoda), David B. Carlini, Kimberly S. Reece, John Graves

VIMS Articles

Phylogenetic analysis conducted on a 784-bp fragment of 82 actin gene sequences of 44 coleoid cephalopod taxa, along with results obtained from genomic Southern blot analysis, confirmed the presence of at least three distinct actin loci in coleoids. Actin isoforms were characteri zed through phylogenetic analysis of representative cephalopod sequences from each of the three isoforms, along with translated actin cDNA sequences from a diverse array of metazoan taxa downloaded from GenBank. One of the three isoforms found in cephalopods was closely related to actin sequences expressed in the muscular tissues of other molluscs. A second isoform was most similar …

Prolonged Cyclooxygenase-2 Induction In Neurons And Glia Following Traumatic Brain Injury In The Rat, K I Strauss, M F Barbe, R M Marshall Demarest, R Raghupathi, S Mehta, R K Narayan

Rowan-Virtua School of Osteopathic Medicine Faculty Scholarship

Cyclooxygenase-2 (COX2) is a primary inflammatory mediator that converts arachidonic acid into precursors of vasoactive prostaglandins, producing reactive oxygen species in the process. Under normal conditions COX2 is not detectable, except at low abundance in the brain. This study demonstrates a distinctive pattern of COX2 increases in the brain over time following traumatic brain injury (TBI). Quantitative lysate ribonuclease protection assays indicate acute and sustained increases in COX2 mRNA in two rat models of TBI. In the lateral fluid percussion model, COX2 mRNA is significantly elevated (>twofold, p < 0.05, Dunnett) at 1 day postinjury in the injured cortex and bilaterally in the hippocampus, compared to sham-injured controls. In the lateral cortical impact model (LCI), COX2 mRNA peaks around 6 h postinjury in the ipsilateral cerebral cortex (fivefold induction, p < 0.05, Dunnett) and in the ipsilateral and contralateral hippocampus (two- and six-fold induction, respectively, p < 0.05, Dunnett). Increases are sustained out to 3 days postinjury in the injured cortex in both models. Further analyses use the LCI model to evaluate COX2 induction. Immunoblot analyses confirm increased levels of COX2 protein in the cortex and hippocampus. Profound increases in COX2 protein are observed in the cortex at 1-3 days, that return to sham levels by 7 days postinjury (p < 0.05, Dunnett). The cellular pattern of COX2 induction following TBI has been characterized using immunohistochemistry. COX2-immunoreactivity (-ir) rises acutely (cell numbers and intensity) and remains elevated for several days following TBI. Increases in COX2-ir colocalize with neurons (MAP2-ir) and glia (GFAP-ir). Increases in COX2-ir are observed in cerebral cortex and hippocampus, ipsilateral and contralateral to injury as early as 2 h postinjury. Neurons in the ipsilateral parietal, perirhinal and piriform cortex become intensely COX2-ir from 2 h to at least 3 days postinjury. In agreement with the mRNA and immunoblot results, COX2-ir appears greatest in the contralateral hippocampus. Hippocampal COX2-ir progresses from the pyramidal cell layer of the CA1 and CA2 region at 2 h, to the CA3 pyramidal cells and dentate polymorphic and granule cell layers by 24 h postinjury. These increases are distinct from those observed following inflammatory challenge, and correspond to brain areas previously identified with the neurological and cognitive deficits associated with TBI. While COX2 induction following TBI may result in selective beneficial responses, chronic COX2 production may contribute to free radical mediated cellular damage, vascular dysfunction, and alterations in cellular metabolism. These may cause secondary injuries to the brain that promote neuropathology and worsen behavioral outcome.

Social Structure Of A Polygynous Tent-Making Bat, Cynopterus Sphinx (Megachiroptera), Jay F. Storz, Hari Bhat, Thomas H. Kunz

Jay F. Storz Publications

The social structure of an Old World tent-making bat Cynopterus sphinx (Megachiroptera), was investigated in western India. A combination of census and mark–recapture data over 2 years (1996–98) was used to infer the form of the mating system, compositional stability of social groups and mode of new social group formation. The breeding population of C. sphinx was subdivided into diurnal roosting colonies, each of which contained one to five discrete roosting groups and often one or more solitary bats in adjacent roosts. Bats most frequently roosted in stem tents constructed in the flower/fruit clusters of the kitul palm Caryota urens …

The Occurrence Of The Freshwater Clams, Musculium Partumeium (Say) And Pisidium Casertanum (Poli) (Bivalvia: Sphaeriidae), In The Hawaiian Islands, Albert J. Burky, Carl M. Way, Skippy Hau, M. Eric Benbow

Biology Faculty Publications

Sphaeriid clams were collected from an ancient, continuously cultivated taro pond complex at Ke‘anae Peninsula, Maui. Both species are known for adaptations for temporary pond habitats and could have been initially introduced by the earliest Hawaiian settlers transported with moist taro root stock from other areas of Oceania.

"An Evolutionary No Man’S Land" And "Reply From L. G. Harshman And A. A. Hoffmann", Margarida Matos, André Levy, Henrique Teotónio, Michael R. Rose, Lawrence G. Harshman, Ary A. Hoffmann

Lawrence G. Harshman Publications

The gap between evolutionary studies in laboratory versus natural populations is a persistent problem.

A Radiation Hybrid Map Of The Cat Genome: Implications For Comparative Mapping, William J. Murphy, Shan Sun, Zhang-Qun Chen, Naoya Yuhki, Deborah Hirschmann, Marilyn Menotti-Raymond, Stephen J. O'Brien

Biology Faculty Articles

Ordered gene maps of mammalian species are becoming increasingly valued in assigning gene variants to function in human and animal models, as well as recapitulating the natural history of genome organization. To extend this power to the domestic cat, a radiation hybrid (RH) map of the cat was constructed integrating 424 Type I-coding genes with 176 microsatellite markers, providing coverage over all 20 feline chromosomes. Alignment of parallel RH maps of human and cat reveal 100 conserved segments ordered (CSOs) between the species, nearly three times the number observed with reciprocal chromosome painting analyses. The observed number is equivalent to …

Unusual Polymorphisms In Human Immunodeficiency Virus Type 1 Associated With Nonprogressive Infection, Louis Alexander, Emma Weiskopf, Thomas C. Greenough, Nathan C. Gaddis, Marcy C. Auerbach, Michael H. Malim, Stephen J. O'Brien, Bruce D. Walker, John L. Sullivan, Ronald C. Desrosiers

Biology Faculty Articles

Factors accounting for long-term nonprogression may include infection with an attenuated strain of human immunodeficiency virus type 1 (HIV-1), genetic polymorphisms in the host, and virus-specific immune responses. In this study, we examined eight individuals with nonprogressing or slowly progressing HIV-1 infection, none of whom were homozygous for host-specific polymorphisms (CCR5-Δ32, CCR2-64I, and SDF-1-3^'A) which have been associated with slower disease progression. HIV-1 was recovered from seven of the eight, and recovered virus was used for sequencing the full-length HIV-1 genome; full-length HIV-1 genome sequences from the eighth were determined following amplification of viral …

Electrically Mediated Plasmid Dna Delivery To Hepatocellular Carcinomas In Vivo, L. Heller, M. J. Jaroszeski, D. Coppola, C. Pottinger, R. Gilbert, Richard Heller

Bioelectrics Publications

Gene therapy by direct delivery of plasmid DNA has several advantages over viral gene transfer, but plasmid delivery is less efficient. In vivo electroporation has been used to enhance delivery of chemotherapeutic agents to tumors in both animal and human studies. Recently, this delivery technique has been extended to large molecules such as plasmid DNA. Here, the successful delivery of plasmids encoding reporter genes to rat hepatocellular carcinomas by in vivo electroporation is demonstrated.

Test Of Intron Predictions Reveals Novel Splice Sites, Alternatively Spliced Mrnas And New Introns In Meiotically Regulated Genes Of Yeast, Carrie Davis, Leslie Grate, Marc Spingola, Manuel Ares

Biology Department Faculty Works

Correct identification of all introns is necessary to discern the protein-coding potential of a eukaryotic genome. The existence of most of the spliceosomal introns predicted in the genome of Saccharomyces cerevisiae remains unsupported by molecular evidence. We tested the intron predictions for 87 introns predicted to be present in non-ribosomal protein genes, more than a third of all known or suspected introns in the yeast genome. Evidence supporting 61 of these predictions was obtained, 20 predicted intron sequences were not spliced and six predictions identified an intron-containing region but failed to specify the correct splice sites, yielding a successful prediction …

New World Nine-Primaried Oscine Relationships: Constructing A Molecular Framework, John Klicka, Kevin P. Johnson, Scott M. Lanyon

Ornithology Program (HRC)

Historically, a paucity of comparative morphological characters has led to much debate regarding relationships within and among the major lineages of New World nine-primaried oscines. More recently, DNA-DNA hybridization studies have provided novel and testable hypotheses of relationships, although no consensus has been reached. For 40 songbird taxa, we obtained 1,929 base pairs (bp) of DNA sequence from the mitochondrial cytochrome-b (894 bp) and NADH dehydrogenase subunit 2 (1,035 bp) genes. Phylogenetic analyses confirm the monophyly of this assemblage as traditionally defined. The lineages delineated historically on morphological grounds are retained; finches (Fringillinae) are sister to a well-supported clade …

Dispersion And Site Fidelity In A Tent-Roosting Population Of The Short-Nosed Fruit Bat (Cynopterus Sphinx) In Southern India, Jay F. Storz, J. Balasingh, P. Thiruchenthil Nathan, K. Emmanuel, Thomas H. Kunz

Jay F. Storz Publications

Patterns of dispersion and site fidelity were investigated in a tent-roosting population of the short-nosed fruit bat, Cynopterus sphinx (Megachiroptera), in southern India. A local population of C. sphinx occupied diurnal roosts in a variable subset of 45 stem tents constructed within the dense foliage of mast trees (Polyalthia longifolia). Individually marked tent-roosting bats were visually censused over the course of a 38-d interval spanning the postpartum oestrus period. On any given day, 33.3-85.7% (mean = 60.8%, SD = 14.2) of adult males roosted singly, with the remainder holding harems of 1-10 breeding females (mean = 3.01, SD …

Oxidative Stress Resistance: A Robust Correlated Response To Selection In Extended Longevity Lines Of Drosophila Melanogaster?, Lawrence G. Harshman, Beth A. Haberer

Lawrence G. Harshman Publications

Stress resistance is associated with longevity in Drosophila melanogaster and other model organisms used for genetic research. The present study tests for oxidative stress resistance in one set of lines selected for late-life reproduction and extended longevity. Both females and males from the selected lines were appreciably more resistant to oxidative stress than were flies from the control lines. A relative increase in oxidative stress resistance is a correlated response to selection in this laboratory selection experiment. Increased oxidative stress resistance appears to be a relatively robust correlated response to laboratory selection for late-life reproduction and extended longevity.

Cell Cycle-Dependent Binding Of Yeast Heat Shock Factor To Nucleosomes, Christina Bourgeois Venturi, Alexander M. Erkine, David S. Gross

Scholarship and Professional Work – COPHS

In the nucleus, transcription factors must contend with the presence of chromatin in order to gain access to their cognate regulatory sequences. As most nuclear DNA is assembled into nucleosomes, activators must either invade a stable, preassembled nucleosome or preempt the formation of nucleosomes on newly replicated DNA, which is transiently free of histones. We have investigated the mechanism by which heat shock factor (HSF) binds to target nucleosomal heat shock elements (HSEs), using as our model a dinucleosomal heat shock promoter (hsp82-ΔHSE1). We find that activated HSF cannot bind a stable, sequence-positioned nucleosome in G₁-arrested …

The Skn7 Response Regulator Of Saccharomyces Cerevisiae Interacts With Hsf1 In Vivo And Is Required For The Induction Of Heat Shock Genes By Oxidative Stress, Desmond C. Raitt, Anthony L. Johnson, Alexander M. Erkine, Kozo Makino, Brian Morgan, David S. Gross, Leland H. Johnston

Scholarship and Professional Work – COPHS

The Skn7 response regulator has previously been shown to play a role in the induction of stress-responsive genes in yeast, e.g., in the induction of the thioredoxin gene in response to hydrogen peroxide. The yeast Heat Shock Factor, Hsf1, is central to the induction of another set of stress-inducible genes, namely the heat shock genes. These two regulatory trans-activators, Hsf1 and Skn7, share certain structural homologies, particularly in their DNA-binding domains and the presence of adjacent regions of coiled-coil structure, which are known to mediate protein–protein interactions. Here, we provide evidence that Hsf1 and Skn7 interact in vitro and …

A Simple Method For Generating Full Length Cdna From Low Abundance Partial Genomic Clones, Yongxin Wang, Joseph M. Fugaro, Fauzia Siddiq, Chandra Mouli V. Goparaju, Fulvio Lonardo, Anil Wali, John F. Lechner, Harvey I. Pass

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

PCR amplification of target molecules involves sequence specific primers that flank the region to be amplified. While this technique is generally routine, its applicability may not be sufficient to generate a desired target molecule from two separate regions involving intron /exon boundaries. For these situations, the generation of full-length complementary DNAs from two partial genomic clones becomes necessary for the family of low abundance genes.

Results

The first approach we used for the isolation of full-length cDNA from two known genomic clones of Hox genes was based on fusion PCR. Here we describe a simple and efficient method …

Identification Of The Transactivation Domain Of The Transcription Factor Sox-2 And An Associated Co-Activator, Tamara K. Nowling, Lance R. Johnson, Matthew S. Wiebe, Angie Rizzino

Nebraska Center for Virology: Faculty Publications

The importance of interactions between Sox and POU transcription factors in the regulation of gene expression is becoming increasingly apparent. Recently, many examples of the involvement of Sox-POU partnerships in transcription have been discovered, including a partnership between Sox-2 and Oct-3. Little is known about the mechanisms by which these factors modulate transcription. To better understand the molecular interactions involved, we mapped the location of the transactivation do-main of Sox-2. This was done in the context of its interaction with Oct-3, as well as its ability to transactivate as a fusion protein linked to the DNA-binding domain of Gal4. Both …

Recent Progress In Psychiatric Genetics—Some Hope But No Hype, Scott F. Stoltenberg, Margit Burmeister

Department of Psychology: Faculty Publications

The reputation of the field of psychiatric genetics has recently become tarnished in the view of many human geneticists. Too many linked loci were claimed and withdrawn, too many association studies published and not confirmed, and, more recently, too many new and different chromosomal regions have been implicated for the same disorder. Here, we summarize recent trends, focusing on research that moves away from traditional linkage studies. Some promising strategies include psychopharmacogenetics and consideration of endophenotypes such as neurophysiological and behavioral markers in addition to the clinical diagnosis. Utilization of rapid and automated methods for scoring genetic variants in large-scale …

Rapid Communication: Porcine Vitamin D-25-Hydroxylase Maps To Chromosome 5, Daniel C. Ciobanu, R. K. Raven, M. F. Rothschild, N. H. Bell

Department of Animal Science: Faculty Publications

Locus Name. Vitamin D-25-hydroxylase.

Species. Sus scrofa.

Locus Symbol. CYP25.

Source of Primers. PCR primers were designed from the published sequence for porcine vitamin D-25-hydroxylase (CYP25) cDNA (GenBank accession no. Y16417). The position of the forward and reverse primers corresponds to 506 to 530 bp and 830 to 854 bp, respectively, of the cDNA. The primer pair amplified a 348-bp fragment of the cDNA and an 862-bp fragment from the pig genomic DNA. The 862-bp fragment (Gen- Bank accession no. AF284378) was subcloned into a pNoTA vector and identity was confirmed by sequencing. Genomic sequences from 1 to 74, …

Comparison Of Plasma Fsh Concentration In Boars And Gilts From Lines Selected For Ovulation Rate And Embryonal Survival, And Litter Size And Estimation Of (Co)Variance Components For Fsh And Ovulation Rate, J. D. Cassady, Rodger K. Dennison, J. J. Ford

Department of Animal Science: Faculty Publications

The objective of this research was to determine whether plasma concentration of FSH was genetically correlated with ovulation rate and thus was a useful trait for indirect selection. Blood samples were collected from 619 animals from five lines of pigs. Line I was selected for increased index of ovulation rate and embryonal survival, and Line C was its randomly selected control. Pigs sampled from Lines I and C were from generations 12 and 13. Pigs from three additional lines that were derived from eighth-generation pigs of Lines I and C also were used. These lines were Line C2, a randomly …

Haemochromatosis Mutation Analysis In A Normal Irish Population, Fergus Ryan, Joseph Vaughan

Articles

187 Irish blood donors were examined for the presence of C282Y and H63D missense mutations using a multiplex PCR and restriction digest protocol. The allele frequency for C282Y was found to be 12.3% which is higher than reported in other studies and concurs that this mutation is of Celtic origin.