Genetics and Genomics Commons^™

Molecular Characterization And Expression Analysis Of Putative Rpk1 And Putative Clv1 In Malaysian Indica Rice (Oryza Sativa L.) Cv.Mr219 And Malaysian Wild Rice (Oryza Rufipogon) Irgc105491., Law Yee Song

Student Works (2010-2019)

Two candidate yield-related genes, putative Receptor-like Protein Kinase 1 (putative RPK1) and putative CLAVATA1 Receptor Kinase (putative CLV1), were selected for molecular characterization and expression analysis. This was carried out using in four rice lines comprising the parents and two BC₂F₇ lines from a local rice breeding program, with Malaysian rice variety, Oryza sativa ssp. indica cv. MR219 as recurrent parent, and Oryza rufipogon (IRGC105491) as donor parent. Approximately 2.8 kb and 2.6 kb putative RPK1 cDNA nucleotide sequences were obtained from Oryza rufipogon and Oryza sativa ssp. indica cv. MR219 respectively. However, only 402 …

Characterization Of Microrna172 And Its Potential Target In Oil Palm (Elaeis Guineensis Jaq.), Fatemeh Mehrpooyan

Student Works (2010-2019)

Oil palm (Elaeis guineensis) is the world’s major perennial monocot crop and contributes highly to the economies of South East Asia. As the oil palm fruit is the source of the main economic output for this crop, floral development is a critical process related to oil palm productivity. miR172 has been shown to be an important regulator of the AP2 family transcription factors involved in floral timing in a number of plants and belongs to a group of highly conserved miRNA that can be identified based on sequence homology. Six oil palm miR172 paralogs, representing two distinct mature miR172 sequences, …

Phylogeographical Patterns Of Neobalanocarpus Heimii (Dipterocarpaceae) For The Evolutionary History And Chain Of Custody Certification., Tnah Lee Hong

Student Works (2010-2019)

Tectonic movement and climatic oscillations during the Cenozoic have had dramatic effect on the biota of the tropical rain forest. This study aims to reveal the phylogeography and evolutionary history of a Peninsular Malaysian endemic tropical timber, Neobalanocarpus heimii (Dipterocarpaceae), based on chloroplast DNA (cpDNA) variation. Fifteen haplotypes were identified from 10 intraspecific variable sites of five non-coding cpDNA regions: trnL intron, trnS-trnG spacer, trnG intron, trnK intron and psbK-trnS spacer. Two major genealogical cpDNA lineages of N. heimii were elucidated: a widespread southern and a northern region. The species is predicted to survive in multiple refugia during climatic oscillation: …

Enterobacterial Repetitive Intergenic Consensus-Pcr (Eric-Pcr) Analysis Among Raw Vegetables Isolates Of Campylobacter Jejuni., Tuan Suraya Tuan Soh

Student Works (2010-2019)

A total of 20 (n=20) raw vegetable isolates of Campylobacter jejuni were examined for the presence of virulence genes and genetic diversity using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) analysis. All C. jejuni isolates were obtained from the laboratory of Food Science and Biotechnology, University Putra Malaysia, Serdang, Selangor. When specific PCR amplification were determined, all raw vegetables isolates of Campylobacter jejuni contained cadF and ceuE genes. Whereas, 12 (12/20) isolates were positive cdtB and 6 (6/20) were positive cdtC. None of the C. jejuni isolates was cdtA positive. Using ERIC-PCR analysis, all 20 isolates of C. jejuni were …

Biolistic Transformation Of Oil Palm Using The Phosphomannose Isomerase (Pmi) Gene As The Positive Selectable Marker., Bahariah Bohari

Student Works (2010-2019)

The selectable marker system based on the Escherichia coli phosphomannose isomerase (pmi) gene was adapted for genetic transformation of oil palm. This system makes use of the pmi gene that encodes phosphomannose isomerase, which converts mannose-6-phosphate to fructose-6-phosphate and uses mannose as the selection agent. This is to anticipate the future requirement of using non-antibiotic resistance genes for the commercialization of transgenic oil palm. The use of antibiotic or herbicide-based selection systems has caused much public concern due to inadequate knowledge of the agents’ impact on the environment and on human health. In this study, four transformation vectors, namely pMI3, …

Genotype And Allele Frequencies Of Nine Str Loci In Sikhs From Malaysia., Tan Chin Teck

Student Works (2010-2019)

Allele frequencies for nine short tandem repeats (STRs) gene loci (CSF1PO, TPOX,TH01, F13A01, FESFPS, vWA, D16S539, D7S820, D13S317) were obtained from analyses of saliva DNA from 109 unrelated healthy Sikh individuals residing in Malaysia. DNA was extracted using phenol/chloroform/isoamly alcohol method. 2ng of DNA for each sample was amplified following the instruction in GenePrint® STR Systems (Promega Corporation) CTT, FFv and SilverSTR III Multiplex kits. PCR products were separated in denaturing polyacrylamide gel electrophoresis and silver stained using Promega DNA Silver Staining System. The results were then calculated and processed to establish a statistical database for Sikh community. Chi-square test …

A Systematic Study Of Fagraea Sensu Lato And A Revision Of The Peninsular Malaysian Taxa., Sugumaran Manickam

Student Works (2010-2019)

systematic study of the Peninsular Malaysian species of Fagraea sensu lato was undertaken. This was done in the light of recent revisionary work done for Borneo that documented 20 new species and demonstrated that previous species concepts for Fagraea were too broad. Parallel to this the distinction of the subgeneric groups recognised as sections, i.e., Cyrtophyllum, Fagraea and Racemosae, were investigated with molecular phylogenetic methods. Representative taxa from the Malay Peninsula and Borneo augmented by sequences from other taxa in the same subtribe and tribe were used in the molecular analysis. Gene sequences from ITS, trnL–F (trnL intron + trnL–F …

Computational Identification Of Micro Rna In Musa Sp Est Sequences., Tan Yew Seong

Student Works (2010-2019)

MicroRNAs (miRNAs) are small single-stranded non-protein-coding RNAs of about 22 nucleotides that play an important role in post-transcriptional gene regulation in animals and plants by targeting mRNAs for direct cleavage of mRNAs or repression of mRNA translation. In this study, a homology search based approach has been used for identifying the novel miRNA genes of Musa sp. using the BLAST program with a total of 42,978 Musa sp. expressed sequence tags (EST) were compared to a total of 959 previously known plant miRNA sequences. Forty-seven candidate miRNA with homology to previously known miRNAs were identified. By using the mFold3.1 program, …

Isolation And Development Of Microsatellite Markers Of Channa Striata., Faiza Omar

Student Works (2010-2019)

Channa striata, locally known as haruan has long been regarded as a valuable edible fish and it is also consumed for its putative effects on wound healing. The main objective of the project was to develop microsatellite markers for use in the study of genetic diversity of the wild Channa striata populations in Malaysia. In this study, seven new microsatellite loci were isolated from Channa striata using a Random Amplified Microsatellite (RAMs) based technique which was an efficient and reliable method. Ten primers were used in population genetic studies of Channa striata.Sixty sets of microsatellite primer pairs were used in …

Agrobacterium-Mediated Transformation Of Cryptocoryne Willisii With Gus And Mgfp 5 Reporter Genes / Wong Sher Ming, Wong Sher Ming

Student Works (2010-2019)

In this study, a simple method using Murashige and Skoog basal medium (1962) supplemented with 6-BA alone was developed for direct multiple shoots induction from shoot base culture of C. willisii. Shoots proliferation and elongation were achieved on a single medium without the need of subculturing. The optimum concentration of 6-BA for direct shoots regeneration of C. willisii was investigated. It was found to be 6 mg/ L. Optimum media was then used for regenerate and propagate of transformed C. willisii. On the other hand, ransformation of C. willisii with Agrobacterium tumefaciens strain LBA4404 harbouring pCAMBIA1304 was performed. Effects on …

Application Of Pcr-Based Serogrouping Of Salmonella Enterica., Mohammed Elbagir Mohammed Elhassan Nori

Student Works (2010-2019)

Serotyping is the most fundamental approach in epidemiological surveillance and outbreak investigations for Salmonella. Many multiplex PCR assays have been reported for serotyping Salmonella based on O and H antigens. The aim of this study was to apply and improve the approach in differentiation and serotyping Salmonella enterica strains by combination of sequential multiplex PCRs targeting the O, H and Vi antigens of Salmonella, and this achieved by working on 126 strains. All the strains were purified on BSA and produced black colonies of Salmonella. The strains were confirmed by PCR amplification gene that targets the hilA. 122 of 126 …

Light Whole Genome Sequence For Snp Discovery Across Domestic Cat Breeds, James C. Mullikin, Nancy F. Hansen, Lei Shen, Heather Ebling, William F. Donahue, Wei Tao, David J. Saranga, Adrianne Brand, Marc J. Rubenfield, Alice C. Young, Pedro Cruz, Carlos Driscoll, Victor David, Samer W. K. Al-Murrani, Mary F. Locniskar, Mitchell S. Abrahamsen, Stephen J. O'Brien, Douglas R. Smith, Jeffrey A. Brockman

Biology Faculty Articles

Background

The domestic cat has offered enormous genomic potential in the veterinary description of over 250 hereditary disease models as well as the occurrence of several deadly feline viruses (feline leukemia virus -- FeLV, feline coronavirus -- FECV, feline immunodeficiency virus - FIV) that are homologues to human scourges (cancer, SARS, and AIDS respectively). However, to realize this bio-medical potential, a high density single nucleotide polymorphism (SNP) map is required in order to accomplish disease and phenotype association discovery.

Description

To remedy this, we generated 3,178,297 paired fosmid-end Sanger sequence reads from seven cats, and combined these data with the …

Molecular Evolution And The Role Of Oxidative Stress In The Expansion And Functional Diversification Of Cytosolic Glutathione Transferases, Rute R. Da Fonseca, Warren E. Johnson, Stephen J. O'Brien, Vitor Vasconcelos, Agostinho Antunes

Biology Faculty Articles

Background: Cytosolic glutathione transferases (cGST) are a large group of ubiquitous enzymes involved in detoxification and are well known for their undesired side effects during chemotherapy. In this work we have performed thorough phylogenetic analyses to understand the various aspects of the evolution and functional diversification of cGSTs. Furthermore, we assessed plausible correlations between gene duplication and substrate specificity of gene paralogs in humans and selected species, notably in mammalian enzymes and their natural substrates.

Results: We present a molecular phylogeny of cytosolic GSTs that shows that several classes of cGSTs are more ubiquitous and thus have an …

Accounting For Multiple Comparisons In A Genome-Wide Association Study (Gwas), Randall C. Johnson, George W. Nelson, Jennifer L. Troyer, J. A. Lautenberger, Bailey Kessing, Cheryl Winkler, Stephen J. O'Brien

Biology Faculty Articles

Background

As we enter an era when testing millions of SNPs in a single gene association study will become the standard, consideration of multiple comparisons is an essential part of determining statistical significance. Bonferroni adjustments can be made but are conservative due to the preponderance of linkage disequilibrium (LD) between genetic markers, and permutation testing is not always a viable option. Three major classes of corrections have been proposed to correct the dependent nature of genetic data in Bonferroni adjustments: permutation testing and related alternatives, principal components analysis (PCA), and analysis of blocks of LD across the genome. We consider …

Increased Perfusion And Angiogenesis In A Hindlimb Ischemia Model With Plasmid Fgf-2 Delivered By Noninvasive Electroporation, B. Ferraro, Y. L. Cruz, M. Baldwin, D. Coppola, R. Heller

Bioelectrics Publications

Gene therapy approaches delivering fibroblast growth factor-2 (FGF-2) have shown promise as a potential treatment for increasing blood flow to ischemic limbs. Currently, effective noninvasive techniques to deliver plasmids encoding genes of therapeutic interest, such as FGF-2, are limited. We sought to determine if intradermal injection of plasmid DNA encoding FGF-2 (pFGF) followed by noninvasive cutaneous electroporation (pFGFE+) could increase blood flow and angiogenesis in a rat model of hindlimb ischemia. pFGFE+ or control treatments were administered on postoperative day 0. Compared to injection of pFGF alone (pFGFE-), delivery of pFGFE+ significantly increased FGF-2 expression for 10 days. Further, the …

Ecology And Genetics Of Philodina Megalotrocha (Rotifera, Bdelloidea) From Chihuahuan Desert Populations, Lina Kamel Hamdan

Open Access Theses & Dissertations

The Chihuahuan Desert is a unique region with high biodiversity. There is a high degree endemism of freshwater biota, but relatively few studies have been focused on zooplankton. Bdelloid rotifers are one component of these ecosystems and often dominate very temporary habitats. Bdelloids are usually assumed to be cosmopolitan species due to their potentially high dispersal rates. Distribution records of the bdelloid P. megalotrocha extend from North America to New Zealand. However, little is known about its ecology or genetics. The aims of this study are to: 1) determine whether the distribution of P. megalotrocha in the Chihuahuan Desert is …

Analysing The Effects Of Loss Of Sin3 In Drosophila Melanogaster, Aishwarya Swaminathan

Wayne State University Dissertations

Sin3A has been previously shown to be an essential gene for Drosophila viability and is implicated in the regulation of cell cycle. In this study, we show that SIN3 is not only required for embryonic viability but also for post-embryonic development. Genetic analysis suggests that the different isoforms of SIN3 may regulate unique sets of genes during development. The developmental lethality occurring due to ubiquitous knock down of SIN3 is hypothesized to be to the result of defects in cell proliferation. Conditional knock down of SIN3 in the wing discs results in a curly wing phenotype in the adult fly. …

Tracking Profiles Of Genomic Instability In Spontaneous Transformation And Tumorigenesis, Lesley Lawrenson

Wayne State University Dissertations

The dominant paradigm for cancer research focuses on the identification of specific genes for cancer causation and for the discovery of therapeutic targets. Alternatively, the current data emphasize the significance of karyotype heterogeneity in cancer progression over specific gene-based causes of cancer. Variability of a magnitude significant to shift cell populations from homogeneous diploid cells to a mosaic of structural and numerical chromosome alterations reflects the characteristic low-fidelity genome transfer of cancer cell populations. This transition marks the departure from micro-evolutionary gene-level change to macro-evolutionary change that facilitates the generation of many unique karyotypes within a cell population. Considering cancer …

Determination Of The Essential Functions Of A Conserved Cyclin, Cyclin Y, In Drosophila, Dongmei Liu

Wayne State University Dissertations

The Drosophila gene CG14939 encodes a member of a highly conserved family of cyclins, the Y type cyclins, which have not been functionally characterized in any organism. Here I report the generation and phenotypic characterization of a null mutant of CG14939, which we rename Cyclin Y (CycY). I show that the null mutant, CycY^E8, is homozygous lethal with most mutant animals arresting during pupal development. The mutant exhibits delayed larval growth and major developmental defects during metamorphosis. Heat shock-induced expression of CycY at different times during development resulted in variable levels of rescue, the timing …

The Role Of Ledgf/P75 In Transcriptional Regulation, Jeffrey Ryan Kugelman

Open Access Theses & Dissertations

The Lens Epithelial Derived Growth Factor p75 (LEDGF/p75) is a chromatin bound protein whose cellular function is not yet clearly known. A role in transcriptional regulation had been previously proposed based on its interaction with the basal transcriptional machinery and on its effects on the expression of genes involved in the cellular response to environmental stresses. To further elucidate the function of LEDGF/p75, we conducted a global and unbiased evaluation of the role of this protein in gene expression. To that aim, we performed a microarray analysis of cellular gene expression in cells that are severely depleted of LEDGF/p75. To …

Genbank, Dennis A. Benson, Ilene Karasch-Mizrachi, David J. Lipman, James Ostell, Eric W. Sayers

Harold W. Manter Laboratory of Parasitology: Library Materials

GenBank(R) is a comprehensive database that contains publicly available nucleotide sequences for more than 380,000 organisms named at the genus level or lower, obtained primarily through submissions from individual laboratories and batch submissions from large-scale sequencing projects, including whole genome shotgun (WGS) and environmental sampling projects. Most submissions are made using the web-based BankIt or standalone Sequin programs, and accession numbers are assigned by GenBank staff upon receipt. Daily data exchange with the European Nucleotide Archive (ENA) and the DNA Data Bank of Japan (DDBJ) ensures worldwide coverage. GenBank is accessible through the NCBI Entrez retrieval system that integrates data …

Building The Genomic Base-Layer Of The Oral “Omic” World, Forsyth Metagenomic Support Consortium, Jacques Izard

Department of Food Science and Technology: Faculty Publications

With the shift of molecular technologies directed toward the understanding of greater biological complexity of the oral cavity, a knowledge gap was created by the lack of genomic data from the diverse oral microorganisms. To facilitate and enable the interpretation of metagenomic, transcriptomic, and proteomic data generated or soon to be generated from oral biofilms, we are providing reference genomic information from phylogenetically diverse oral bacterial isolates. This work, initiated by the National Institute of Dental and Craniofacial Research as an isolated effort, is now part of the Human Microbiome Project. The goal of this effort is the public release …

Neuroaids In Africa, Kevin Robertson, Jeff Liner, James Hakim, Jean-Louis Sankalé, Igor Grant, Scott Letendre, David Clifford, Amadou Gallo Diop, Assan Jaye, Georgette Kanmogne, Alfred Njamnshi, T. Dianne Langford, Tufa Gemechu Weyessa, Charles Wood, Mwanza Banda, Mina Hosseinipour, Ned Sacktor, Noeline Nakasuja, Paul Bangirana, Robert Paul, John Joska, Joseph Wong, Michael Boivin, Penny Holding, Betsy Kammerer, Annelies Van Rie, Prudence Ive, Avindra Nath, Kathy Lawler, Clement Adebamowo, Walter Royal Iii, Jeymohan Joseph

Nebraska Center for Virology: Faculty Publications

In July 2009, the Center for Mental Health Research on AIDS at the National Institute of Mental Health organized and supported the meeting “NeuroAIDS in Africa.” This meeting was held in Cape Town, South Africa, and was affiliated with the 5th IAS Conference on HIV Pathogenesis, Treatment and Prevention. Presentations began with an overview of the epidemiology of HIV in sub-Saharan Africa, the molecular epidemiology of HIV, HIV-associated neurocognitive disorders (HANDs), and HAND treatment. These introductory talks were followed by presentations on HAND research and clinical care in Botswana, Cameroon, Ethiopia, The Gambia, Kenya, Malawi, Nigeria, Senegal, South Africa, Uganda, …

Functional Properties Of The Hiv-1 Subtype C Envelope Glycoprotein Associated With Mother-To-Child Transmission, Hong Zhang, Marzena Rola, John T. West, Damien C. Tully, Piotr Kubis, Jun He, Chipepo Kankasa, Charles Wood

Nebraska Center for Virology: Faculty Publications

Understanding the properties of viruses capable of establishing infection during perinatal transmission of HIV-1 is critical for designing effective means of limiting transmission. We previously demonstrated that the newly transmitted viruses (in infant) were more fit in growth, as imparted by their envelope glycoproteins, than those in their corresponding mothers. Here, we further characterized the viral envelope glycoproteins from six mother-infant transmission pairs and determined whether any specific envelope functions correlate with HIV-1 subtype C perinatal transmission. We found that most newly transmitted viruses were less susceptible to neutralization by their maternal plasma compared to contemporaneous maternal viruses. However, the …

Chronology And Evolution Of The Hiv-1 Subtype C Epidemic In Ethiopia, Damien C. Tully, Charles Wood

Nebraska Center for Virology: Faculty Publications

Objective—To reconstruct the onset date and evolutionary history of the HIV-1 subtype C epidemic in Ethiopia - one of the earliest recorded subtype C epidemics in the world.

Design—HIV-1 C env sequences with a known sampling year isolated from HIV-1 positive patients from Ethiopia between 1984 and 2003.

Methods—Evolutionary parameters including origin and demographic growth patterns were estimated using a Bayesian coalescent-based approach under either strict or relaxed molecular clock models.

Results—Bayesian evolutionary analysis indicated a most recent common ancestor date of 1965 with three distinct epidemic growth phases. Regression analysis of root-to-tip distances revealed a highly similar estimate for …

Enhancement Of Autophagy During Lytic Replication By The Kaposi’S Sarcoma-Associated Herpesvirus Replication And Transcription Activator, Hui-Ju Wen, Zhilong Yang, You Zhou, Charles Wood

Nebraska Center for Virology: Faculty Publications

Autophagy is one of two major degradation systems in eukaryotic cells. The degradation mechanism of autophagy is required to maintain the balance between the biosynthetic and catabolic processes and also contributes to defense against invading pathogens. Recent studies suggest that a number of viruses can evade or subvert the host cell autophagic pathway to enhance their own replication. Here, we investigated the effect of autophagy on the KSHV (Kaposi’s sarcoma-associated herpesvirus) life cycle. We found that the inhibition of autophagy reduces KSHV lytic reactivation from latency, and an enhancement of autophagy can be detected during KSHV lytic replication. In addition, …

Molecular Basis Of A Novel Adaptation To Hypoxic-Hypercapnia In A Strictly Fossorial Mole, Kevin L. Campbell, Jay F. Storz, Anthony V. Signore, Hideaki Moriyama, Kenneth C. Catania, Alexander P. Payson, Joseph Bonaventura, Jörg Stetefeld, Roy E. Weber

Jay F. Storz Publications

Background: Elevated blood O₂ affinity enhances survival at low O₂ pressures, and is perhaps the best known and most broadly accepted evolutionary adjustment of terrestrial vertebrates to environmental hypoxia. This phenotype arises by increasing the intrinsic O₂ affinity of the hemoglobin (Hb) molecule, by decreasing the intracellular concentration of allosteric effectors (e.g., 2,3-diphosphoglycerate; DPG), or by suppressing the sensitivity of Hb to these physiological cofactors.

Results: Here we report that strictly fossorial eastern moles (Scalopus aquaticus) have evolved a low O₂ affinity, DPG-insensitive Hb - contrary to expectations for a mammalian species that is …

Phenotypic Plasticity And Genetic Adaptation To High-Altitude Hypoxia In Vertebrates, Jay F. Storz, Graham R. Scott, Zachary A. Cheviron

Jay F. Storz Publications

High-altitude environments provide ideal testing grounds for investigations of mechanism and process in physiological adaptation. In vertebrates, much of our understanding of the acclimatization response to high-altitude hypoxia derives from studies of animal species that are native to lowland environments. Such studies can indicate whether phenotypic plasticity will generally facilitate or impede adaptation to high altitude. Here, we review general mechanisms of physiological acclimatization and genetic adaptation to high-altitude hypoxia in birds and mammals. We evaluate whether the acclimatization response to environmental hypoxia can be regarded generally as a mechanism of adaptive phenotypic plasticity, or whether it might sometimes represent …

Genetic Differences In Hemoglobin Function Between Highland And Lowland Deer Mice, Jay F. Storz, Amy M. Runck, Hideaki Moriyama, Roy E. Weber, Angela Fago

Jay F. Storz Publications

In high-altitude vertebrates, adaptive changes in blood–O₂ affinity may be mediated by modifications of hemoglobin (Hb) structure that affect intrinsic O₂ affinity and/or responsiveness to allosteric effectors that modulate Hb–O₂ affinity. This mode of genotypic specialization is considered typical of mammalian species that are high-altitude natives. Here we investigated genetically based differences in Hb–O₂ affinity between highland and lowland populations of the deer mouse (Peromyscus maniculatus), a generalist species that has the broadest altitudinal distribution of any North American mammal. The results of a combined genetic and proteomic analysis revealed that deer mice harbor …

Genome3d: A Viewer-Model Framework For Integrating And Visualizing Multi-Scale Epigenomic Information Within A Three-Dimensional Genome, Thomas M. Asbury, Matt Mitman, Jijun Tang, W. Jim Zheng

Faculty Publications

Background
New technologies are enabling the measurement of many types of genomic and epigenomic information at scales ranging from the atomic to nuclear. Much of this new data is increasingly structural in nature, and is often difficult to coordinate with other data sets. There is a legitimate need for integrating and visualizing these disparate data sets to reveal structural relationships not apparent when looking at these data in isolation.

Results
We have applied object-oriented technology to develop a downloadable visualization tool, Genome3D, for integrating and displaying epigenomic data within a prescribed three-dimensional physical model of the human genome. In order …