Genetics and Genomics Commons^™

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 Too many linked loci were claimed and withdrawn, too many association studies published and not confirmed, and, more recently, too many new and different chromosomal regions have been implicated for the same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 …

Studying The Expression Of A Novel Microsatellite Mrna In Chironomous Riparius, Mina Ranzurmal

Seton Hall University Dissertations and Theses (ETDs)

The expression of a unique mRNA containing a microsatellite sequence was studied in C. riparius using differential display. First, the rnicrosatellite fragment was sequenced using Sanger's Sequencing method. The microsatellite cDNA contained 11 "ACA" repeats, 12 "AC" repeats, and 5 "CTC" repeats. Second, the affects of cadmium and heat shock on rnicrosatellite mRNA expression were studied. In cadmium experiments, larvae were exposed to low cadmium concentrations (0, 0.04, 0.4, and 4.0 mM) for 24 and 48 h and high cadmium concentrations (0, 1.0, 2.0, 4.0, 8.0, and 16.0 mM) for 48 h. The effects of cadmium on fed larvae at …

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 …

Whole Genome Sequencing Of L.Monocytogenes And L.Innocua : A Basis For Comparative Genomics Of The Genus Listeria, Rachel Purcell

Theses

Listeria monocytogenes is a gram-positive, food-borne human and animal pathogen. L.innocua is a non-pathogenic species the most closely related to L. monocytogenes. Comparative genomics between a pathogenic and a closely related non-pathogenic species is a powerful approach for identifying genes or regions on the chromosome, which might be involved in virulence. Therefore whole genome sequencing of these two organisms was undertaken. The strategy used was based on a random approach combined with end-sequencing of a medium insert library for the finishing phase. Based on the available sequence data for these two species an in silico analysis for the presence of …

Application Of Genetic Markers To Provide Species Identification And Define Stock Structure: Analyses Of Selected Marine Fishes Of The Mid -Atlantic Bight, Jan F. Cordes

Dissertations, Theses, and Masters Projects

Molecular markers and techniques were employed to develop a genetic key for the forensic identification of 16 species of Chesapeake Bay sportfishes and to investigate the stock structure of one of those species, the weakfish Cynoscion regalis. Regions within the ATP synthetase 6 (ATPase 6), Cytochrome b , cytochrome c oxidase I, NADH dehydrogenase 4 (ND4), and 12S/16S ribosomal RNA mitochondrial genes were amplified using the polymerase chain reaction (PCR) and digested with a bank of restriction endonucleases to find a genetic marker that exhibited complete interspecific differentiation and low intraspecific variation. Complete separation of all sixteen species was accomplished …

A Molecular Phylogeny Of The Sparidae (Perciformes: Percoidei), Thomas M. Orrell

Dissertations, Theses, and Masters Projects

The Sparidae are a diverse group of over 110 marine species whose putative six subfamilies have been defined primarily on the basis of dentition and feeding type. Monophyly of the subfamilies has not been tested, nor have the phylogenetic relationships of all sparid genera been hypothesized. I used mitochondrial DNA sequences from the complete cytochrome b gene (1140 bp) and the partial 16S gene (621 bp) in independent and combined analyses to test the monophyly of the Sparidae, elucidate the inter-relationships of the 33 recognized genera of the Sparidae, test the validity of the six subfamilies of the Sparidae, and …

Transcriptional Regulation Of Chicken Apolipoprotein A-I Gene Expression, Wen Lin Kumfert

Theses and Dissertations in Biomedical Sciences

Deregulation of a set of critical cellular genes has long been speculated as a cause of the oncogenic transformation induced by v-Jun oncoprotein. In the past several years, extensive efforts have been made to identify such genes. Several target genes have been identified that are specifically associated with the v-Jun induced transformation phenotype in chicken embryo fibroblasts (CEF).

We have undertaken the objective to identify and characterize the genes that become deregulated in response to, or as a consequence of, Jun-induced transformation. By exploiting the difference in oncogenic potential between v-Jun and c-Jun in CEF, the chicken apolipoprotein A-I (apoA-I) …

Analysis Of The Population Genetics Of The Han And Hui Of Liaoning Province, Peoples Republic Of China, Michael L. Black

Theses: Doctorates and Masters

Throughout recorded Chinese history, regions of the country populated by persons of non-Han ancestry often fluctuated significantly in population numbers and in their political and commercial influence. However, at all times they were considered as important contributors to the nation. Many of these peoples had moved from their homelands, settled in China and had intermarried with Han Chinese. Over the generations they became accepted as fully-fledged Chinese citizens although, in many instances, they retained their traditional customs and religious practices, and frequently their own language. The Hui Muslims are a good example of this process of integration, and today they …

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.

Analysis Of Genetic Factors Affecting High Density Lipoprotein Levels And Apoall Levels In C57 B1/6j And Fvb/N Wildtype Mice Using A Qtl Mapping Approach, Judith Barrios

Seton Hall University Dissertations and Theses (ETDs)

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Scientific Investigation Of The Plant Pathogen Agrobacterium Tumefaciens, Casey M. Flanagan, Christopher B. Crowell Ii, Bryan P. Markelz

Honors Theses

A. tumefaciensis a plant pathogen and causative agent of Crown Gall Disease, an ailment that results in the development of tumors in plants. It is a unique organism in that it is the only known non-viral pathogen that transfers its DNA into the host cell. Consequently, the majority of the research conducted on A. tumefaciensinvestigates its mechanism of infection. However, A. tumefaciensis also rare to the bacterial world in that its genome is comprised of two chromosomes: one circular and one linear. Most other bacteria only have one circular chromosome. This area of research has been grossly overlooked and therefore …