Browsing by Subject "comparative genomics"
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Item Construction of a minimal tiling path across the euchromatic arms of sorghum chromosome 3 and comparative analysis with the rice chromosome 1 pseudomolecule(2009-05-15) Zhou, BinUsing rice chromosome 1 pseudomolecule as a reference, a minimal tiling path for the euchromatic arms of sorghum chromosome 3 was constructed, in which 23 contigs contain an estimated 57.56 Mb of DNA. A total of 409 EST-STS markers and 255 genetic markers have been mapped onto the euchromatic arms providing excellent integration of the genetic and physical maps. A total of 21 contigs containing 9 ESTSTS and 35 genetic markers have been constructed across the heterochromatin block of sorghum chromosome 3 which comprise 16.57 Mb of DNA. Macrocolinearity between sorghum chromosome 3 and rice chromosome 1 was examined based on the mapped EST-STS markers. Approximately 85% of the EST-STS markers were colinear between these two homeologous chromosomes. Estimates of recombination were also determined, which indicates the existence of recombination cold and hot spots. Microcolinearity between sorghum chromosome 3 and rice chromosome 1 was examined at two different levels. In one case, overlapping sorghum BAC pools orthologous to a 5.1 Mb region of rice chromosome 1 were constructed and sequence skimmed. Alignment of the sorghum skim sequences to the TIGR rice gene models revealed ~62% colinearity between the two orthologous regions. In addition, colinearity between sorghum chromosome 3 and rice chromosome 5 was detected within this region which is likely due to the segmental homology between rice chromosome 1 and rice chromosome 5. Microcolinearity between sorghum and rice was also examined by comparing 2 fully sequenced sorghum chromosome 3 BAC clones to the orthologous region of rice chromosome 1. In this analysis, ~65% colinearity was detected for sorghum BAC 82G24 and ~59% colinearity was detected for sorghum BAC 181g10. Microcolinearity was largely confined to gene coding regions and sequences of exons displayed the highest percent identities. Small-scale gene rearrangements were also detected. Finally, RT-PCR analysis was carried out between a set of colinear and non-colinear genes from sorghum and rice to determine whether the loss of colinearity between orthologous genes resulted in a change in transcriptional regulation. No direct link between loss of colinearity and expression pattern was detected in these experiments.Item Molecular and Genetic Analysis of Adaptive Evolution in the Rare Serpentine Endemic, Caulanthus amplexicaulis var. barbarae (J. Howell) Munz(2011-10-21) Burrell, Anna MildredIn the interest of understanding the genetic basis of adaption to environment, we developed F2 lines from an F1 interspecific cross between the rare serpentine endemic, Caulanthus amplexicaulis var. barbarae and the non-serpentine Caulanthus amplexicaulis var. amplexicaulis. Using genomic DNA from Caulanthus amplexicaulis var. barbarae, we developed a suite of microsatellite markers. In addition, we developed gene specific markers for genes known in Arabidopsis to be ecologically important. Our suite of markers was used to genotype 186 F2 plants, the basis for our F2 linkage map. In order to further resolve evolutionary relationships among related taxa, we constructed a molecular phylogeny for 52 taxa within the related genera Caulanthus, Guillenia, Sibaropsis, Streptanthella, and Streptanthus, using the sequences from the ribosomal ITS region and two chloroplast regions. To create a useful system to enable comparative genomics within the related taxa of the ecologically and morphologically diverse Streptanthoid Complex, we demonstrated that our molecular tools are portable across a large group of ecologically significant taxa. To use the significant genomic resources available in Arabidopsis, we constructed a collinear comparative map of Caulanthus and the model plant Arabidopsis thaliana based on ancestral linkage blocks with the Brassicaceae family. This comparative map acted as a guide for candidate gene selection in the mapping of sepal color. We identified a region of MYB transcription factors in an orthologous region of Arabidopsis. Sequence data from Caulanthus amplexicaulis var. barbarae and Caulanthus amplexicaulis var. amplexicaulis in this MYB region showed significant sequence divergence between the two taxa. To determine the genetic basis for the tolerance of high concentrations of magnesium in Caulanthus amplexicaulis var. barbarae, we phenotyped multiple individuals from 88 F2:3 families under two nutrient treatments, differing in the ratio of calcium to magnesium. Through QTL analysis, using our F2 linkage map as a framework for the analysis, we identified one major effect QTL on Caulanthus Linkage Group 8 and another QTL on Caulanthus Linkage Group 3. We identified candidate genes for the QTLs using our collinear comparative map to Arabidopsis.Item Organization of the class I region of the bovine major histocompatibility complex (BoLA) and the characterization of a class I frameshift deletion (BoLA-Adel) prevalent in feral bovids(Texas A&M University, 2006-04-12) Ramlachan, NicoleThe major histocompatibility complex (MHC) is a genomic region containing genes of immunomodulatory importance. MHC class I genes encode cell-surface glycoproteins that present peptides to circulating T cells, playing a key role in recognition of self and non-self. Studies of MHC loci in vertebrates have examined levels of polymorphism and molecular evolutionary processes generating diversity. The bovine MHC (BoLA) has been associated with disease susceptibility, resistance and progression. To delineate mechanisms by which MHC class I genes evolved to function optimally in a species like cattle, it is necessary to study genomic organization of BoLA to define gene content, and investigate characteristics of expressed class I molecules. This study describes development of a physical map of BoLA class I region derived from screening two BAC libraries, isolating positive clones and confirming gene content, order and chromosomal location through PCR, novel BAC end sequencing techniques, and selected BAC shotgun cloning and/or sequencing and FISH analysis. To date, this is the most complete ordered BAC array encompassing the BoLA class I region from the class III boundary to the extended class I region. Characterization of a frameshift allele exhibiting trans-species polymorphism in Bos and Bison by flow cytometry, real-time RT-PCR, 1D and 2D gel analysis is also described. This frameshift allele encodes an early termination signal within the antigen recognition site (ARS) of exon 3 of the BoLA BSA-Adel class I gene predicting a truncated class I protein that is soluble. An ability to assess MHC diversity in populations and provision of animals with defined MHC haplotypes and genetic content for experimental research is necessary in developing a basis upon which to build functional studies to elucidate associations between haplotype and disease in bovids. The BoLA class I region is immunologically important for disease association studies in an economically important species. This study provides knowledge of gene content and organization within the class I MHC region in cattle, providing a template for more detailed analysis and elucidation of complex disease associations through functional genomics and comparative analysis, as well as evolution of the MHC in bovids to optimize a population??s immune response.