Thesis (M.S.)--Brigham Young University. Dept. of Computer Science, 2006. / Includes bibliographical references (p. 39-41).

This dissertation explores the genetic basis of gene expression in Fundulus heteroclitus by focusing on the role of the environment and its effects on gene expression and by making direct estimates of heritability using cDNA microarrays. The second chapter describes the utility of F. heteroclitus cDNA microarrays for studies of F. heteroclitus which seek to understand the genetic variation in gene expression. Measurements of mRNA fluorescence and concentration as well as differences in sample preparation and sampling of blood from a single individual over time demonstrate that F. heteroclitus cDNA microarrays are quantitative, reproducible and consistent. The third chapter examines the effect of the environment and genetic factors on the variation of gene expression. F. heteroclitus cDNA microarrays are used to determine whether a genetic component of gene expression can describe the variation in gene expression between inbred and outbred individuals from the same population. The results show that variation in mRNA expression is related to the genetic variation among individuals within a group. While chapter three reveals that there is a genetic component of variation in gene expression, the percentage of genes that are significantly heritable was not known. In the fourth chapter, the heritability of the variation in gene expression is estimated to determine the genetic basis of gene expression in F1 individuals from natural, outbred populations of F. heteroclitus. The data presented in chapter 4 are the first to formally estimate the genetic component of gene expression in F. heteroclitus. The estimates of heritability range from 0.25 to 0.86 depending on the estimation method with approximately 6.5% of genes having significant heritability. The results presented in this dissertation support the concept that genetic variation affects variation in mRNA expression among natural populations of F. heteroclitus. Natural, heritable variation in gene expression is important for understanding evolutionary adaptation and the role of natural selection in evolutionary processes.

The overall aim of the studies described herein was to evaluate genetic variation in cattle fertility traits for development of genetic and genomic predictors in breeding strategies. Results from these experiments suggest that improvements in fertility through genetic selection are a possible approach to increase reproductive efficiency. Experiment 1 evaluated the development of genetic parameters associated with multiple ovulation and embryo transfer schemes in an attempt to assist producers in identifying animals with greater genetic merit for these protocols. This study confirmed that genetic selection of donors or sires appears to be a potential approach to improve efficiency of MOET procedures. Although low heritability would slow the progress, results shown in this work suggest that genetic improvement in fertility by selection for embryo transfer traits is possible. Experiment 2 evaluated fertility traits in Argentinean Holstein cattle in order to develop fertility genetic predictors for utilization in breeding strategies. The dollar fertility index ($F) included age to first calving (AFC) as a measure of initial reproductive performance and calving interval (CI) as an indicator of conception rate and success of early insemination. Values for $F ranged from -$76.6 to $139.4 in the current Holstein population. Results indicated substantial variation in fertility traits, suggesting that genetic selection would be highly effective in improving fertility.

Thesis (Ph. D.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

In the budding yeast Saccharomyces cerevisiae, the G1-specific cyclin-dependent kinases (Cdks) Cln1-, Cln2-Cdc28 and Pcl1-, Pcl2-Pho85 are essential for ensuring that DNA replication and cell division are properly linked to cell polarity and bud morphogenesis. However, like most genes in S. cerevisiae, individual cyclin genes are not required for viability, and the phenotypes associated with deletion of any single cyclin gene tend to be subtle. My goal was to dissect the cellular roles of the G1 cyclins by systematically identifying their genetic interactions. To do this, I conducted Synthetic Genetic Array (SGA) screens using strains deleted for different combinations of cyclin genes. The results of screens with strains deleted for the G1 cyclin pairs, CLN1, CLN2, or PCL1, PCL2, confirmed a role for these cyclins in cell polarity regulation and identified novel G1 Cdk substrates, which I examined in more detail. One cell polarity regulator that showed an interesting pattern of genetic interactions with G1 cyclins was BNI1, which encodes a yeast formin protein. Overexpression of BNI1 caused an Synthetic Dosage Lethal interaction in the absence of both G1 cyclin pairs while its deletion caused synthetic lethality specifically in the absence of PCL1, PCL2. Consistent with these genetic interactions, phosphorylation of Bni1 was partially dependent on CLN1, CLN2. It has been proposed that Bni1 is regulated by intramolecular interactions. In an effort to discover how phosphorylation might affect Bni1 function, I developed assays to test for intramolecular interactions. In my experiments I found no evidence that Bni1 is regulated by intramolecular binding, as was proposed from parallels with its mouse homolog mDia1. I also found that deletion of BNI4, which encodes an adaptor protein that targets several proteins to the bud neck, results in severe growth defects in the absence of the Cdc28 cyclins Cln1 and Cln2, and overexpression of BNI4 was toxic in yeast cells lacking the Pho85 cyclins Pcl1 and Pcl2. I discovered that Bni4 was phosphorylated by Pcl1- and Pcl2-Pho85 in vitro and that phosphorylation of Bni4 was dependent on PCL1 and PCL2 in vivo. Further analysis showed that phosphorylation of Bni4 by Pcl-Pho85 is necessary for its localization to bud neck, and the bud neck structure can be disrupted by overexpressing BNI4 in pcl1pcl2 mutant cells. I propose that if Bni4 cannot be regulated by phosphorylation, it may titrate away an essential component that resides at the bud neck, thus causing catastrophic morphogenesis defects. The relationship between G1 Cdk activity and the polarity regulator Bni4 serves as a bridge to link the cell cycle machine to the regulation of cell.

Genes modulating innate immunity in mammals are generally considered the first line of defense with respect to invading pathogens and therefore it has become important to characterize naturally occurring genetic variation, and subsequently determine whether this variation is likely to be benign, beneficial, or detrimental to the host. Relevant to this study, the mammalian Toll-like receptor proteins (TLR), encoded by members of the TLR gene family, have the capacity to recognize a wide variety of pathogen ligands, and mutations within these genes have been shown to influence disease susceptibility or resistance within mammalian species. Two studies which sought to determine the frequency and distribution of naturally occurring genetic variation within the bovine and equine TLR genes revealed a large number of discrete point mutations, which were subsequently used to reconstruct haplotypes for each investigated gene across a large number of samples. Detailed analyses of haplotypes provided evidence for extensive haplotype sharing among specialized breeds, subspecies, and even divergent species. Classical and new tests of selection provided evidence for significant deviations from a strictly neutral model of molecular evolution for both cattle as well as equids, with some of the same TLR genes deviating from a strictly neutral model among divergent species. As a first step toward determining whether naturally occurring bovine TLR variation is likely to be benign, beneficial, or detrimental, we tested validated variation from bovine TLR genes capable of recognizing components of Mycobacteria for associations with Mycobacterium avium subspecies paratuberculosis (MAP) infection in dairy cattle, and found several SNPs that were nominally associated with disease status, thereby providing evidence for small-effect loci potentially influencing risk for differential susceptibility to Johne's disease.

In the budding yeast Saccharomyces cerevisiae, the G1-specific cyclin-dependent kinases (Cdks) Cln1-, Cln2-Cdc28 and Pcl1-, Pcl2-Pho85 are essential for ensuring that DNA replication and cell division are properly linked to cell polarity and bud morphogenesis. However, like most genes in S. cerevisiae, individual cyclin genes are not required for viability, and the phenotypes associated with deletion of any single cyclin gene tend to be subtle. My goal was to dissect the cellular roles of the G1 cyclins by systematically identifying their genetic interactions. To do this, I conducted Synthetic Genetic Array (SGA) screens using strains deleted for different combinations of cyclin genes. The results of screens with strains deleted for the G1 cyclin pairs, CLN1, CLN2, or PCL1, PCL2, confirmed a role for these cyclins in cell polarity regulation and identified novel G1 Cdk substrates, which I examined in more detail. One cell polarity regulator that showed an interesting pattern of genetic interactions with G1 cyclins was BNI1, which encodes a yeast formin protein. Overexpression of BNI1 caused an Synthetic Dosage Lethal interaction in the absence of both G1 cyclin pairs while its deletion caused synthetic lethality specifically in the absence of PCL1, PCL2. Consistent with these genetic interactions, phosphorylation of Bni1 was partially dependent on CLN1, CLN2. It has been proposed that Bni1 is regulated by intramolecular interactions. In an effort to discover how phosphorylation might affect Bni1 function, I developed assays to test for intramolecular interactions. In my experiments I found no evidence that Bni1 is regulated by intramolecular binding, as was proposed from parallels with its mouse homolog mDia1. I also found that deletion of BNI4, which encodes an adaptor protein that targets several proteins to the bud neck, results in severe growth defects in the absence of the Cdc28 cyclins Cln1 and Cln2, and overexpression of BNI4 was toxic in yeast cells lacking the Pho85 cyclins Pcl1 and Pcl2. I discovered that Bni4 was phosphorylated by Pcl1- and Pcl2-Pho85 in vitro and that phosphorylation of Bni4 was dependent on PCL1 and PCL2 in vivo. Further analysis showed that phosphorylation of Bni4 by Pcl-Pho85 is necessary for its localization to bud neck, and the bud neck structure can be disrupted by overexpressing BNI4 in pcl1pcl2 mutant cells. I propose that if Bni4 cannot be regulated by phosphorylation, it may titrate away an essential component that resides at the bud neck, thus causing catastrophic morphogenesis defects. The relationship between G1 Cdk activity and the polarity regulator Bni4 serves as a bridge to link the cell cycle machine to the regulation of cell.

Infertility in dairy cattle remains a major economic loss to dairy producers. Identifying dairy cattle with superior genetic potential for improved fertility would increase dairy farm profitability. Dairy heifers were classified into two groups based upon services per conception (SPC); those animals with a single SPC were determined to be highly fertile and animals with greater than or equal to 4 SPC were classified as sub-fertile. Whole genome association analysis was performed on 20 individual heifers from each group utilizing a 777K highly density (HD) single nucleotide polymorphism (SNP) chip. Genomic data were evaluated utilizing PLINK, a whole genome association analysis toolset, and 570,620 SNP were available for analysis with a total of 39 samples being analyzed. Forty-four SNP were determined to be associated with fertility classification (P <= 0.00001) and were located on Bos taurus chromosome (BTA) 2, 4, 9, 19, and 26. The SNP and ranges between SNP were analyzed using BLAST-Like Alignment Tool (BLAT); SNP were associated with 5 candidate genes for reproduction. The SNP on BTA 2 were located within the region coding for the non-imprinted Prader-Willi/Angelman syndrome 2 (NIPA2) gene, which is involved in gestational magnesium transport. Also on BTA 2, SNP were identified within the region encoding for cytoplasmic fragile X mental retardation 1 (FMR1) interaction protein 1 (CYFIP1). The CYFIP1 gene is involved with the functionality of FMR1 and has been linked to premature ovarian failure in humans. Additionally, 3 SNP on BTA 9 were located near monofunctional C1-tetrahydrofolate synthase (MTHFD1L), which has been linked to neural tube defects during gestation in humans A difference in allele frequency was observed between the two groups for SNP located on BTA19 in proximity to two genes, zinc finger 18 (ZNF18) and mitogen activated protein kinase 4 (MAP2K4). The ZNF18 motif and MAP2K4 were found to be involved in heart development of the early embryo and associated with toll-like receptors (TLR) involved in gonadotropin releasing hormone (GnRH) signaling, respectively. The involvement of one or all of these genes may further explain reduced fertility in dairy cattle.

Thesis (M.S. in App. Bio.)--School of Biology, Georgia Institute of Technology, 2004. Directed by Roger M. Wartell. / Includes bibliographical references (leaves 34-37).

Thesis (Ph. D.)--University of Chicago, Committee on Developmental Biology, December 2003. / Includes bibliographical references. Also available on the Internet.