Candidate gene mapping: approach, methods and significance

Abstract Candidate gene is a gene with known or assumed function that may affect genetic control of a trait and thus, can be considered a 'candidate gene' for this trait. The Candidate gene associates a gene to its phenotypic trait. These quantitative traits responsible may be biomedical, economical, and even evolutionary important studies. The traditional candidate gene identification is tedious due to limited information of molecular marker and, also lack of computational tools and software. However, digital candidate gene approach makes candidate gene identification reliable and rapid due to available literature database and gene ontology database. The Candidate gene mapping is successfully conducted with the identification of molecular marker, linkage map construction and Quantitative trait locus mapping. The candidate gene approach is important for determination of associated genetic variant with phenotype

QTL global meta-analysis: are trait determining genes clustered?

Background: A key open question in biology is if genes are physically clustered with respect to their known functions or phenotypic effects. This is of particular interest for Quantitative Trait Loci (QTL) where a QTL region could contain a number of genes that contribute to the trait being measured. Results: We observed a significant increase in gene density within QTL regions compared to non-QTL regions and/or the entire bovine genome. By grouping QTL from the Bovine QTL Viewer database into 8 categories of non-redundant regions, we have been able to analyze gene density and gene function distribution, based on Gene Ontology (GO) with relation to their location within QTL regions, outside of QTL regions and across the entire bovine genome. We identified a number of GO terms that were significantly over represented within particular QTL categories. Furthermore, select GO terms expected to be associated with the QTL category based on common biological knowledge have also proved to be significantly over represented in QTL regions. Conclusion: Our analysis provides evidence of over represented GO terms in QTL regions. This increased GO term density indicates possible clustering of gene functions within QTL regions of the bovine genome. Genes with similar functions may be grouped in specific locales and could be contributing to QTL traits. Moreover, we have identified over-represented GO terminology that from a biological standpoint, makes sense with respect to QTL category type.Hanni Salih and David L Adelso

Integrating linkage and radiation hybrid mapping data for bovine chromosome 15

BACKGROUND: Bovine chromosome (BTA) 15 contains a quantitative trait loci (QTL) for meat tenderness, as well as several breaks in synteny with human chromosome (HSA) 11. Both linkage and radiation hybrid (RH) maps of BTA 15 are available, but the linkage map lacks gene-specific markers needed to identify genes underlying the QTL, and the gene-rich RH map lacks associations with marker genotypes needed to define the QTL. Integrating the maps will provide information to further explore the QTL as well as refine the comparative map between BTA 15 and HSA 11. A recently developed approach to integrating linkage and RH maps uses both linkage and RH data to resolve a consensus marker order, rather than aligning independently constructed maps. Automated map construction procedures employing this maximum-likelihood approach were developed to integrate BTA RH and linkage data, and establish comparative positions of BTA 15 markers with HSA 11 homologs. RESULTS: The integrated BTA 15 map represents 145 markers; 42 shared by both data sets, 36 unique to the linkage data and 67 unique to RH data. Sequence alignment yielded comparative positions for 77 bovine markers with homologs on HSA 11. The map covers approximately 32% of HSA 11 sequence in five segments of conserved synteny, another 15% of HSA 11 is shared with BTA 29. Bovine and human order are consistent in portions of the syntenic segments, but some rearrangement is apparent. Comparative positions of gene markers near the meat tenderness QTL indicate the region includes separate segments of HSA 11. The two microsatellite markers flanking the QTL peak are between defined syntenic segments. CONCLUSIONS: Combining data to construct an integrated map not only consolidates information from different sources onto a single map, but information contributed from each data set increases the accuracy of the map. Comparison of bovine maps with well annotated human sequence can provide useful information about genes near mapped bovine markers, but bovine gene order may be different than human. Procedures to connect genetic and physical mapping data, build integrated maps for livestock species, and connect those maps to more fully annotated sequence can be automated, facilitating the maintenance of up-to-date maps, and providing a valuable tool to further explore genetic variation in livestock

Identification of genomic regions that contribute to wet carcass syndrome in sheep

Wet carcass syndrome (WCS), which is observed predominantly in sheep, affects carcass quality negatively. After slaughter the carcass appears to be ‘wet’ with a subcutaneous accumulation of watery fluid. Not all animals in a contemporary group are afflicted, and experimental attempts to induce WCS have been unsuccessful. The reported prevalence of WCS in Dorper and Dorper crosses gave rise to the hypothesis that it may have a genetic basis. Therefore, the primary objective of this investigation was to test this hypothesis using a high-density SNP assay to search loci that may predispose sheep to WCS. Muscle samples from 43 afflicted and 41 unafflicted sheep were collected from slaughterhouses in the province of Northern Cape, South Africa, and in southern Namibia. Tests against candidate genes proved uninformative, as did runs of homozygosity. Potential associations between WCS and an autosomal genetic marker were investigated further in a case-control genome-wide association study. Separate analyses for each sex were motivated because single nucleotide polymorphisms (SNPs) on the X chromosome suggested quantitative trait loci. These analyses revealed significant associations between SNP and WCS in males, but not in females. Three SNPs that reached genome-wide significance in males are in strong linkage disequilibrium with the Duchenne muscular dystrophy, 5-hydroxytryptamine receptor 2C, and Teneurin transmembrane protein 1 genes. These genes are identified as positional candidate genes, and the Duchenne muscular dystrophy, 5-hydroxytryptamine receptor 2C genes have biological effects that have been documented in other species, making them plausible functional candidate genes for WCS in sheep.Keywords: association analysis, Dorper, sheep carcass, single nucleotide polymorphisms, X chromosom

Mining an Ostrinia nubilalis midgut expressed sequence tag (EST) library for candidate genes and single nucleotide polymorphisms (SNPs)

Genes expressed in lepidopteran midgut tissues are involved in digestion and Bacillus thuringiensis (Bt) toxin resistance traits. Five hundred and thirty five unique transcripts were annotated from 1745 high quality O. nubilalis larval midgut expressed sequence tags (ESTs). Full-length cDNA sequence of 12 putative serine proteinase genes and 3 partial O. nubilalis aminopeptidase N protein genes, apn1, apn3, and apn4, were obtained, and genes may have roles in plant feeding and Bt toxin resistance traits of Ostrinia larvae. The EST library was not normalized and insert frequencies reflect transcript levels under the initial treatment conditions and redundancy of inserts from highly expressed transcripts allowed prediction of putative single nucleotide polymorphisms (SNPs). Ten di-, tri- or tetranucleotide repeat unit microsatellite loci were identified, and minisatellite repeats were observed within the C-termini of two encoded serine proteinases. Molecular markers showed polymorphism at 28 SNP loci and one microsatellite locus, and Mendelian inheritance indicated that markers were applicable to genome mapping applications. This O. nubilalis larval midgut EST collection is a resource for gene discovery, expression information, and allelic variation for use in genetic marker development

Design and analysis of genetical genomics studies and their potential applications in livestock research

Quantitative Trait Loci (QTL) mapping has been widely used to identify genetic loci attributable to the variation observed in complex traits. In recent years, gene expression phenotypes have emerged as a new type of quantitative trait for which QTL can be mapped. Locating sequence variation that has an effect on gene expression (eQTL) is thought to be a promising way to elucidate the genetic architecture of quantitative traits. This thesis explores a number of methodological aspects of eQTL mapping (also known as “genetical genomics”) and considers some practical strategies for applying this approach to livestock populations. One of the exciting prospects of genetical genomics is that the combination of expression studies with fine mapping of functional trait loci can guide the reconstruction of gene networks. The thesis begins with an analysis in which correlations between gene expression and meat quality traits in pigs are investigated in relation to a pork meat quality QTL previously identified. The influence on power due to factors including sample size and records of matched subjects is discussed. An efficient experimental design for two-colour microarrays is then put forward, and it is shown to be an effective use of microarrays for mapping additive eQTL in outbred crosses under simulation. However, designs optimised for detecting both additive and dominance eQTL are found to be less effective. Data collected from livestock populations usually have a pedigreed structure. Many family-based association mapping methods are rather computationally intensive, hence are time-consuming when analysing very large numbers of traits. The application of a novel family-based association method is demonstrated; it is shown to be fast, accurate and flexible for genetical genomics. Furthermore, the results show that multiple testing correction alone is not sufficient to control type I errors in genetical genomics and that careful data filtering is essential. While it is important to limit false positives, it is desirable not to miss many true signals. A multi-trait analysis based on grouping of functionally related genes is devised to detect some of the signals overlooked by a univariate analysis. Using an inbred rat dataset, 13 loci are identified with significant linkage to gene sets of various functions defined by Gene Ontology. Applying this method to livestock species is possible, but the current level of annotations is a limiting factor. Finally, the thesis concludes with some current opinions on the development of genetical genomics and its impact on livestock genetics research

Analysis of host and herpesvirus interactions using bioinformatics.

Bioinformatics methods have become central to analysing and organising the sequence data continually produced by new and existing sequencing projects. The field of bioinformatics covers both the static aspects of organising and presenting these raw data, by compiling existing knowledge into accessible databases, ontologies, and libraries; and the more dynamic aspects of knowledge discovery informatics for interpreting and mining existing data. The aim of this thesis is to utilise such methods to analyse the herpesvirus-host relationship. In Chapter 2 comparative host and herpesvirus genome analysis is used to compare the sequences of all currently sequenced herpesvirus open reading frames to the conceptually translated human genome with the aim of identifying herpesvirus-human (host) sequence homologues. Collating in one search all currently known host homologues provides the first complete assessment of herpesvirus-host homologues. This search identified 55 previously identified herpesvirus-host homologues, and 4 previously unknown herpesvirus-host homologues. The work performed in Chapter 2 highlighted the need for consistent annotation of genomes and gene products to allow greater comparative genomics. It is not feasible to manually curate large numbers of genes whose relationships to each other are not immediately clear. Therefore, Chapters 3 and 4 focus upon the use of the Gene Ontology; a resource that is made publicly available for the purpose of annotating gene products with unified vocabulary derived from a structured directed acyclic graph. The Gene Ontology was extended to allow host-pathogen interaction annotation by a) adding 187 new terms relating specifically to virus function and structure (Chapter 3), and b) using both the new and existing terms to annotate the entire Human Herpesvirus 1 genome using references from the available literature (Chapter 4). Finally, Chapter 5 examines the utility of the Gene Ontology when analysing such large-scale host and herpesvirus gene expression datasets as produced experimentally by DNA microarray studies. Using such automated annotation methods a cluster of 12 proteins were identified that increase mitochondrial function in HUVEC cells 24 hours post HCMV infection. A cluster of nine proteins that function in the MAPK pathway were also identified using the Gene Ontology that provide evidence for HCMV inhibition of the MAPK pathway