Dissertations / Theses on the topic 'Scat DNA'

Conservation translocations are increasingly used worldwide to prevent extinctions and support ecological restoration projects. These translocations involve a lot of uncertainty, particularly when species are introduced to ecosystems where they have not previously coexisted. Many historical translocations in Australia and globally have failed due to insufficient baseline data and inadequate post-translocation monitoring. Contemporary translocations can aim to improve translocation outcomes by increasing baseline data collection and developing robust post-release monitoring for translocated species. The development of environmental DNA (eDNA) techniques has facilitated the collection of information from environments with minimal disturbance to species. The advancement of such passive monitoring techniques has allowed for increased ability to monitor and study cryptic and rare species. The Shark Bay rufous hare-wallaby (Lagorchestes hirsutus bernieri) and the banded hare-wallaby (Lagostrophus fasciatus) were introduced to Dirk Hartog Island (DHI) from founder populations on the Bernier and Dorre islands in Shark Bay, Western Australia, as part of the Dirk Hartog Island National Park Ecological Restoration Program. Once found across large areas of south-west and central Australia, the populations on the Bernier and Dorre islands are the last natural populations of the two species due to habitat loss and predation post-European colonisation. As the rufous hare-wallaby and banded hare-wallaby have not been previously recorded on DHI, there was uncertainty regarding their successful establishment on the island and their potential interactions with each other and other species involved in the restoration program. The diets of rufous hare-wallabies and banded hare-wallabies were investigated through the DNA metabarcoding of scat samples from three islands in Shark Bay, Western Australia. This researched aimed to assess dietary overlap and the potential for resource competition between the two hare-wallaby species. Additionally, the difference in diet between the founder populations to their post-translocation diets on DHI were analysed as an indication of dietary flexibility and adaptability to environmental changes. The diets of both species of hare-wallaby were found to be broad, containing taxa from multiple plant families including invasive weeds. On DHI there was a high degree of overlap in the diets of the two species, indicating a risk of resource competition. The diets of the translocated populations differed significantly from the founder populations, which demonstrated flexible foraging behaviour and signified that rufous hare-wallabies and banded hare-wallabies are excellent candidates for future translocation projects on other islands and within mainland reserves. This thesis is the first study to define and compare the diets of the Shark Bay rufous hare-wallaby and the banded hare-wallaby using scat DNA. The understanding of diet is fundamental to ecology and an essential consideration in restoration projects involving new species interactions. The findings from this research can benefit the fulfilment of translocation success criteria related to species establishment and health targets for the new populations. By demonstrating the dietary breadth and flexibility of Shark Bay’s hare-wallabies, this thesis exemplifies the utility of scat DNA in supporting the conservation of translocated fauna.

Soybean (Glycine max) is an important crop in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were 1) to determine differences between ISSR and RAPD marker systems in detecting genetic variation in soybeans and 2) to identify and characterize accession- diagnostic molecular markers in G. max accessions. Genomic DNAs from 108 G. max accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The levels of polymorphic loci detected with the two marker systems were in general moderate and similar.. Overall, 82% of genetic distance values were above 0.40 based on ISSR analysis. However, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The dendrograms constructed with ISSR data revealed that the South Korean accessions formed an out-group while the RAPD analysis showed that accessions from Sweden were separate from the other 10 gene pools. One variety-diagnostic marker generated with ISSR 5 primer was identified in the accession Kao Chien Tao from China. This marker was cloned, and sequenced. Although RAPD and ISSR marker systems detected similar levels of genetic variability, they target different regions of the soybean genome, resulting in different clustering of the 11 gene pools indicating different genetic relatedness among them. This finding demonstrates the usefulness of both marker systems in assessing diversity and relatedness among Glycine max gene pools.

La voie de signalisation du récepteur des androgènes (AR) est impliquée dans la progression du cancer de la prostate, et il a été montré que des mutations dans ce domaine étaient responsables de l'activation constitutive des gènes placés sous le contrôle des hormones androgènes. Une de ces mutations transforme un résidu thréonine du DBD en alanine (T575A). Des expériences permettant de mesurer l'activité de transcription ont permis à l'équipe du Dr. Ceraline à l'IRCAD de montrer que la mutation T575A induit un changement de spécificité du récepteur. Alors que l'activité de promoteurs placés sous le contrôle d'éléments de réponse spécifique de AR diminue, celle des promoteurs placés sous le contrôle d'éléments non spécifique augmente. Ce changement de spécificité est corrélé à une modification de l'affinité du récepteur pour les éléments de réponse spécifiques et non spécifiques. Afin de comprendre le mécanisme de cette "reprogrammation" à l'échelle moléculaire, l'étude structurale des domaines DBD des récepteurs sauvage et muté a été entreprise par RMN. La comparaison des deux structures en solution a montré que la mutation n'altère pas le repliement du domaine et donc que la différence de reconnaissance des éléments de réponse n'est pas liée directement à la structure tridimensionnelle du domaine. Nous avons ensuite cherché à déterminer si l'altération de la fonction n'était pas due à une différence de dynamique de la chaîne peptidique. Afin d'étudier les mouvements moléculaires le long de la chaîne, des mesures de relaxation hétéronucléaire ont été effectuées et ont montré également une grande similarité dans le comportement dynamique des deux domaines, à l'exception d'une région située dans le premier doigt de zinc à proximité d'une histidine (H570), qui est conservée dans l'ensemble de la famille des domaines DBD des récepteurs nucléaires. Cette différence nous a conduit à mesurer, par RMN, le pKa de cette histidine pour les deux protéines. Nous avons ainsi montré que la mutation T575A induit une diminution de 0,5 unité de pH par rapport à la même histidine dans le domaine sauvage. L'analyse de la structure a permis de montrer que cette différence de pKa est liée à la perte d'une interaction entre le groupe hydroxyle de la thréonine 575 et le cycle imidazole de l'histidine. L'effet de la mutation sur le mécanisme de reconnaissance s'explique donc par un effet indirect dans lequel un acide aminé situé à distance de la région d'interaction modifie la surface électrostatique du domaine DBD. L'effet de la charge positive en position 570 sur la spécificité de reconnaissance de l'élément de réponse a ensuite été étudiée en construisant plusieurs mutants portant ou non une charge à cette position (mutants H570R et H570A). Ces études ont permis de confirmer l'importance de cette charge et l'ensemble de nos travaux fournissent un éclairage inédit sur les mécanismes de reconnaissance de l'ADN par les récepteurs nucléaires. .
The androgen receptor (AR) is a ligand-activated transcriptional factor and a member of the nuclear receptor super family. AR shares a common structural and functional architecture with other members of nuclear receptors. The DNA binding domain of AR (ARDBD) binds to specific response elements as a homodimer. In the clinic, certain mutations in AR are associated with the progression of prostate cancer and have consequences for the treatment of patients with advanced prostate cancer. Previous studies showed that the mutation T575A, locating in the DNA binding domain, enhances the transcriptional activity regulated by full-length AR on promoters containing the non-specific response element compared to the wild type domain does not. These differences prompted us to study the molecular mechanism of ARDBD wild type and the T575A mutant. Structures of ARDBD wild type and T575A mutant revealed high similarity. However, dynamic behavior showed distinct differences between wild type and T575A mutant domains. The protonation state of H570 in ARDBD was found to be differed by the mutation. This loss of charge of H570 results in changes in transcriptional activity of AR. .

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Embora apresente várias limitações, a técnica de RAPD ainda representa a principal ferramenta utilizada no Programa de Melhoramento Genético do Feijoeiro- comum BIOAGRO/UFV. Para contornar os problemas apresentados por esta técnica, a estratégia utilizada por vários pesquisadores tem sido a conversão dos marcadores RAPD em marcadores SCAR e o desenvolvimento de primers microssatélite. Assim, no presente trabalho foram desenvolvidos 5 primers SCAR para mancha-angular, 5 primers para ferrugem e 7 primers para antracnose. Os primers desenvolvidos para mancha-angular derivaram-se dos marcadores OPM02 425 , OPBA16 669 , OPH13 490 e OPH14+AA19 400 e foram mapeados, em acoplamento, a 5,3, 7,1, 5,6 e 10,1 cM, dos respectivos genes de resistência. Os primers desenvolvidos para ferrugem originaram-se dos marcadores OPAE19 890 , OPX11 550 , OPAJ18+AH10 700 , mas apenas o primer sAE19 foi validado. Este foi mapeado, em repulsão, a 1,0 cM do gene de resistência Ur-11. Os primers para antracnose foram desenvolvidos, a partir da seqüência dos fragmentos OPAZ20 940 , OPY20 830 , OPC08 900 , OPZ04 560 , OPB03 1800 , OPZ09 950 , OPH18 830 . Destes, apenas os primers sAZ20, sY20, sC08, sZ04 foram mapeados, em acoplamento, a 7,1, 1,2, 7,8 e 2,9 cM, dos respectivos genes de resistência. Os primers microssatélite foram desenvolvidos, a partir das seqüências de fragmentos de DNA genômico de 3 pequenas bibliotecas enriquecidas. As bibliotecas enriquecidas para as repetições GGC, CCA e AT permitiram a seleção de 79, 172 e 50 clones positivos, respectivamente. Do total de 301 clones que foram identificados, 153 já foram seqüenciados. Observou-se que 40 (26%) clones seqüenciados apresentaram fragmentos de DNA contendo microssatélite, os quais variaram quanto ao tipo, número e tamanho. As seqüências apresentaram repetições de di-, tri-, tetra- e até pentanucleotídeos. Em um mesmo fragmento, foram encontrados até 3 microssatélites. Entre as seqüências analisadas, foram observadas repetições perfeitas, imperfeitas e compostas, variarando entre 12 e 24 pb. Até o momento, foram desenhados e testados 10 pares de primers, sendo um deles originado da seqüência do marcador RAPD OPAZ20 940 . Destes, apenas 5 pares (FCctc001, FCggc001, FCccg001, FCccg002 e FCgca001) amplificaram fragmentos com tamanho esperado e bem definidos. Entretanto, esses primers foram monofórficos, quando testados entre diferentes cultivares andinos e mesoamericanos. Os primers SCAR, desenvolvidos e validados no presente trabalho, foram testados juntamente com 45 pares de primers microssatélite comerciais, entre os genitores de uma população de 154 RIL ́s. Dos primers selecionados, 7 foram mapeados em 10 grupos de um mapa parcial de ligação já existente e 1 permaneceu não-ligado. O mapa de ligação sofreu pequena variação no tamanho (247,8/252 cM) e no número de grupos de ligação (9/10 grupos). Entretanto, pela análise de variância foram constatadas associações significativas desses marcadores com diferentes características quantitativas. As associações mais significativas foram constatadas por meio de análises de regressão stepwise, as quais promoveram alteração no valor de R 2 da regressão múltipla para algumas das características. As características MAT (número de dias até a maturação), VAPLA (número médio de vagens por planta), SEPLA (número médio de sementes por planta) e PRVAG (produção média por vagem), que apresentavam valores de R 2 de 40,14; 28,99; 14,03 e 17,13 , tiveram um aumento para 44,30; 34,53; 21,92 e 26,03, respectivamente. A inclusão do marcador sH13 no GL 07 possibilitou a identificação de um novo QTL para a característica VAPLA. Este marcador também mostrou-se associado ao QTL, anteriormente, descrito para SEPLA. O marcador BM165 foi mapeado no GL 02 e mostrou-se associado a 4 QTLs diferentes, identificados para as características MAT, P100 (peso de 100 sementes), SEPLA e PRVAG.
The RAPD technique represents the major molecular tool for assisting the common bean breeding program of the BIOAGRO/UFV, in spite of its limitations. The conversion of the RAPD markers into SCAR ones, as well as the development of SSR primers are the strategy adopted by several researchers in order to relieve these problems. Therefore, 5 primers SCAR to angular leaf spot, 5 primers to rust and 7 primers to anthracnose were developed. The primers developed to angular leaf spot were derived from the RAPD markers OPM02 425 , OPBA16 669 , OPH13 490 and OPH14+AA19 400 and were mapped, in coupling phase, at 5.3, 7.1, 5.6 and 10.1 cM, from their respective resistance genes. The primers developed to rust were originated from the RAPD markers OPAE19 890 , OPX11 550 , OPAJ18+AH10 700 , although only the primer sAE19 was validated. It was mapped, in repulsion, at 1.0 cM distance from the resistance gene Ur-11. The primers developed for anthracnose were obtained from the fragment sequences of OPAZ20 940 , OPY20 830 , OPC08 900 , OPZ04 560 , OPB03 1800 , OPZ09 950 , OPH18 830 , from wich, only the primers sAZ20, sY20, sC08, sZ04 were validated. They were mapped, in coupling phase, at 7.1, 1.2, 7.8 and 2.9 cM, from their respective resistance genes. The SSR primers were developed from the sequences of the genomic DNA fragments obtained from three small enriched libraries. The libraries were enriched for the repetitions GGC, CCA and AT and made possible the selection of 79, 172 e 50 positive clones, respectively. From the total of 301 positive and identified clones, 153 were sequenced. From these clones, 40 (26%) presented DNA fragments containing microsatellite that showed variations for type, number, and size. The sequences showed repetitions of two, three, four, and five nucleotides. In the same DNA fragment, a total up to three microsatellites were found. Among the analyzed sequences, some perfect, imperfect, and compound repetitions ranging from 12 to 24 bp were found. Until this moment, only 10 pairs of primers were designed and tested; one of them was originated from the RAPD marker OPAZ20 940 . From those, only 5 pairs (FCctc001, FCggc001, FCccg001, FCccg002 e FCgca001) amplified the well-defined fragments with the expected size. However, those primers were monomorphics ones, when tested among different andean and mesoamerican cultivars. The developed and validated SCAR primers were tested together with 45 pairs of commercial SSR primers between the genitors of 154 RIL populations. From the selected primers, 7 were mapped into 10 groups of a partial linkage map already available in the lab, whereas one stayed discoupled. The linkage map was slightly altered in its size (247.8/252 cM) and number of linkage groups (9/10 groups). Variance analysis detected siginificative associations of these markers with different quantitative traits. The most significative associations were detected by stepwise regression analysis that promoted alteration in R 2 values of the multiple regression analysis for some traits. The R 2 values of the traits MAT (the number of days until the bean maturation), VAPLA (the average pod number per plant), SEPLA (the average seed number per plant) and PRVAG (the average yield per pod) increased from 40.14, 28.99, 14.03 and 17.13 to 44.30, 34.53, 21.92 and 26.03, respectively. The marker sH13 included into LG 07 allowed for the identification of a new QTL for the trait VAPLA. This marker also showed to be associated with the QTL previously described for SEPLA. The marker BM165 was mapped in LG 02 and showed to be associated with four different QTLs identified for the traits MAT, P100 (100-seed weight), SEPLA and PRVAG.
Tese importada do Alexandria

Regulation of chromatin by epigenetic modifications is a fundamental step during the control of gene expression in eukaryotic cells. The participation of different factors including histone chaperones, chromatin remodeling complexes and histone-modifying complexes regulate chromatin dynamics and ensure the correct metabolism of transcripts that need to be exported to the cytoplasm. In these lines, post-translational modifications including monoubiquitination of histone H2B (H2Bub1) and methylation of histone H3 represent a well-studied histone cross-talk which is required for chromatin integrity and transcription. Additionally, the transition from H2Bub1 to its deubiquitinated form by Ubp8, the DUB enzyme from SAGA (Spt-Ada-Gcn5-acetyltranferase) co-activator complex, is fundamental to obtain a correct gene expression. In this work, we demonstrate that the histone chaperone Asf1 and the Ran-binding protein Mog1, participate in maintaining correct levels of H2Bub1. We show that Mog1 is required for the trimethylation of histone H3 at lysine 4 (H3K4me3), hence, acting as a modulator of histone cross-talk. Mog1 role into gene expression is also demonstrated by its physical and genetically interaction with transcription factors including SAGA and COMPASS complexes. Indeed, we demonstrate that Mog1 interacts genetically with TREX-2 subunits and affects mRNA export. During this work, we have also focused in understanding the molecular mechanisms surrounding Spinocerebellar Ataxia Type 7 (SCA7) which is a rare disease caused by amino acid glutamine (Q) repeats within the DUBm component, ATXN7. Therefore, our interest has been directed towards the study of new mechanisms that trigger SCA7 such as the DUB activity from SAGA complex, protein-protein interaction networks and metabolic profiles.
La regulación de la cromatina a través de modificaciones epigenéticas es un paso fundamental durante el control de la expresión génica en células eucariotas. La participación de diferentes factores tales como chaperonas de histonas, complejos de remodelación de la cromatina y complejos modificadores de histonas, regulan la dinámica de la cromatina y garantizan el correcto metabolismo de los transcritos que necesitan ser exportados al citoplasma. De esta forma, las modificaciones postraduccionales que incluyen la monoubicuitinación de la histona H2B (H2Bub1) y la metilación de la histona H3 representan un "cross-talk" de histonas la cual es requerida para la integridad de la cromatina y la transcripción. Además, la transición de H2Bub1 a su forma desubicuitinada por Ubp8, la enzima DUB del complejo co-activador SAGA (Spt-Ada-Gcn5-acetiltranferasa), es necesaria para obtener una expresión génica correcta. En este trabajo, se demuestra que la chaperona de histona Asf1 y la proteína de unión a Ran, Mog1, participan en el mantenimiento de los niveles de H2Bub1. Se demuestra que Mog1 es necesaria para la trimetilación de la histona H3 en la lisina 4 (H3K4me3), actuando como un modulador del "cross-talk" de histonas. El papel de Mog1 en la expresión génica también se demuestra por sus interacciones físicas y genéticas con factores de transcripción, incluyendo los complejos SAGA y COMPASS. Además, demostramos que Mog1 interactúa genéticamente con subunidades de TREX-2 y afecta a la exportación de mRNAs. Durante este trabajo, también nos hemos centrado en la comprensión de los mecanismos moleculares que envuelven a la Ataxia Espinocerebelosa Tipo 7 (SCA7), que es una enfermedad rara causada por una repetición de aminoácidos glutamina (Q) dentro del componente del DUBm, ATXN7. Por lo tanto, nuestro interés se ha dirigido hacia el estudio de nuevos mecanismos que desencadenan SCA7, como la actividad DUB del complejo SAGA, las interacciones proteína-proteína y los perfiles metabólicos.
La regulació de la cromatina a través de modificacions epigenètiques és un pas fonamental durant el control de l'expressió gènica en cèl·lules eucariotes. La participació de diferents factors tals com chaperones d'histones, complexos de remodelació de la cromatina i complexos modificadors d'histones, regulen la dinàmica de la cromatina i garanteixen el correcte metabolisme dels transcrits que necessiten ser exportats al citoplasma. D'aquesta forma, les modificacions postraduccionals que inclouen la monoubicuitinació de la histona H2B (H2Bub1) i la metilació de la histona H3 representen un "cross-talk" d'histones la qual és requerida per a la integritat de la cromatina i la transcripció. A més, la transició d'H2Bub1 a la seua forma desubicuitinada per Ubp8, l'enzim DUB del complex co-activador SAGA (Spt-Ada-Gcn5-acetiltranferasa), és necessària per a obtenir una expressió gènica correcta. En aquest treball, es demostra que la chaperona de histona Asf1 i la proteïna d'unió a Ran, Mog1, participen en el manteniment dels nivells d'H2Bub1. Es demostra que Mog1 és necessària per a la trimetilació de la histona H3 en la lisina 4 (H3K4me3), actuant com un modulador del "cross-talk" d'histones. El paper de Mog1 en l'expressió gènica també es demostra per les seues interaccions físiques i genètiques amb factors de transcripció, incloent els complexos SAGA i COMPASS. A més, vam demostrar que Mog1 interactua genèticament amb subunitats de TREX-2 i afecta a l'exportació de mRNA. Durant aquest treball, també ens hem centrat en la comprensió dels mecanismes moleculars que envolten a l'Atàxia Espinocerebelosa Tipus 7 (SCA7), que és una malaltia rara causada per una repetició d'aminoàcids glutamina (Q) dins del component del DUBm, ATXN7. Per tant, el nostre interès s'ha dirigit cap a l'estudi de nous mecanismes que desencadenen SCA7, com l'activitat DUB del complex SAGA, les interacciones proteïna-proteïna i els perfils metabòlics.
Oliete Calvo, P. (2017). Study of the SAGA deubiquitination module: identification of new modulators and its implication on Spinocerebellar Ataxia Type 7 [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/86155
TESIS

No presente estudo investigou-se amostras de plasma com DNA livre circulante (DNA LC) por meio da técnica Varredura Z. Esta é uma técnica eficiente na determinação de parâmetros de diferentes materiais, tais como cristais líquidos, ferrofluidos e compostos biológicos. Esta experiência é realizada através da focalização de um feixe laser de perfil gaussiano numa amostra. Na medida em que a amostra se aproxima do foco da lente, a intensidade do feixe aumenta e alcança seu valor máximo no ponto focal, então diminui para pontos distantes do foco. Na região próxima ao ponto focal se amplificam os fenômenos não-lineares. Recentemente foi demonstrado que níveis elevados de DNA LC no plasma ocorrem com frequência em pacientes com vários tipos de câncer, podendo ser utilizados para discriminar pacientes com malignidade de pessoas saudáveis. As amostras de DNA LC, submetidas ao experimento Varredura Z, forneceram respostas ópticas devido ao fenômeno de lente térmica. Os resultados revelaram que a amplitude de lente térmica das amostras extraídas do plasma de pacientes com malignidade difere daquela de doadores sãos. A técnica Varredura Z se mostrou mais vantajosa em relação a outras biológicas porque revelou uma maior diferença entre os grupos estudados e tem o caráter de detectar mudanças estruturais no DNA LC.
In the present study plasma samples with cell-free DNA were investigated by means of the Z-Scan technique. This is a powerfull technique in determining parameters of different materials, such as liquid crystals, ferrofluids and biological compounds. This experiment is performed by focusing a Gaussian profile laser beam on a sample. As the sample approaches the focus of the lens, the intensity of the beam increases and reaches its maximum value at the focal point, then decreases to points distant from the focus. In the region near the focal point non-linear phenomena are amplified. It has recently been demonstrated that high levels of plasma cell-free DNA occur frequently in patients with various cancers and can be used to discriminate patients with malignancy from healthy donors. The cell-free DNA samples, submitted to the Z-Scan experiment, provided optical responses due to the thermal lens phenomenon. The results revealed that the thermal lens amplitude of samples extracted from the plasma of patients with malignancy differs from that of healthy donors. The Z-Scan technique was more advantageous than other biological ones because it revealed a greater difference between the studied groups and has the character of detecting structural changes in cell-free DNA.

Sunflower (Helianthus annuus L.) high oleic acid content lines differ from conventional sunflower by an increase in oleic acid (C18:1) content of more than 60%. The current sunflower cultivars under production in South Africa are standard sunflower with high levels of linoleic acid (C18:2). The aim of this study was to improve the quality of oil produced by local sunflower germplasm with respect to oleic acid through employing a marker-assisted breeding technique to facilitate and speed up the recovery of the high oleic acid allele into the background of the recurrent parent genome. Eleven sunflower breeding genotypes with high and low oleic acid traits were obtained from the Agricultural Research Council-Grain Crops Institute (ARC-GCI) in Potchefstroom. The breeding genotypes were phenotypically characterised based on their oleic and linoleic acid levels using gas chromatography. Results demonstrated that the average mean of oleic and linoleic acid contents in high oleic acid genotypes were 72% and 17% respectively, while the average mean of oleic acid and linoleic acid contents in wild type lines were 33.5 % and 54 % respectively. These results indicated a perfect negative correlation between the amount of oleic and linoleic acids possessed in high and low oleic acid genotypes (R2 = -99.16%). Sequence characterised amplified region (SCAR) markers were tested to ascertain if any of the ten available dominant FAD2-1 markers was segregating with the high oleic acid allele. Four dominant SCAR markers (FAD2-1F4/R1; FAD2-1F4/R2; FAD2-1F13/R1; FAD2-1F14/R2) were strongly associated with the high oleic acid trait (P< 0.001). With regard to the inheritance of the high oleic acid trait, 143 plants of the F2 segregating population derived from a cross between the high oleic acid parent (AP901-95-3-4-1) and low oleic acid parent (H55-9-2-1-1) were genotyped with the four SCAR markers to determine the genetic state concerning the high oleic acid gene (Ol). Results from a Chi square analysis of the observed frequencies of each dominant FAD2-1 marker locus in 143 F2 individuals indicated that the deviation from the expected ratio of 3:1 (high to low oleic acid) was not statistically significant (P< 0.95) from the observed segregation ratio. These results were consistent with the previous finding that an incomplete dominant gene governs sunflower high oleic acid. A multiplex assay of 78 Simple sequence repeat (SSR) markers was optimised and evaluated on 143 plants of the F2 population to determine suitable SSR markers that can be used in a marker-assisted background selection. Only 14 markers were suitable for marker-assisted background selection based on their high polymorphic information content, allele frequency and maximum allele numbers. In conclusion, this study demonstrated the potential of using SSR and SCAR marker systems as a breeding tool to characterise and speed up the selection process in marker-assisted backcross breeding.
Thesis (MSc (Botany))--North-West University, Potchefstroom Campus, 2013.

This master’s thesis deals with analysis and follow-up suggestion of efficiency improvement on the production line in RACIO s.r.o. First part of thesis is dealing with theoretical preparation and understanding lean production. Second part is dealing with analyzing production process and application of methods lean production. According to the production process analysis at the given production line a new solution of the production process has been suggested to increase production efficiency. This solution has been implemented and compared to the production process before, as well as to the production process after increasing the efficiency.

This master's thesis is engaged in optimisation processes of PUQ (as for Purchasing Quality) department, Robert Bosch České Budějovice. This department is responsible for input control of parts for manufacture as well as check of new parts including their documentation and documentation updates. By optimisation we understand selection of the best variant from group of possibilities.

碩士
國立臺灣大學
農藝學研究所
91
A collection of 25 worldwide representative samples was used to assess the genetic diversity of Southern India type vetiver as well as the development of technique for variety identification. The AFLP marker system was used to study the genetic diversity of Southern India type vetiver. The results indicated the reproducibility of the banding information of the AFLP marker system was 99.5%. Four sets of primers combinations were selected in the analysis and yielded 260 polymorphic bands. One hundred and sixty five polymorphic bands were selected to estimate the genetic distance of the vetiver, which was ranging from 0.09 to 0.64. A cluster analysis with UPGMA methodology of the samples shown the samples from outside India was belong to the group “Sunshine”. The DNA profile information of each one of the Southern India vetiver was established from the AFLP and RAPD polymorphic bands. The three of the four primer combinations in the AFLP analysis were able to establish DNA profile for each one of the samples alone. While the RAPD marker system required information from at least three primers to distinguish the cultivars. Twenty-six and thirty-nine unique bands from RAPD and AFLP, respectively, were identified from the revealed polymorphic bands in the analyses. Ten unique bands were chosen in the development of SCAR marker for the identification of the cultivars in order to improve the accuracy of the variety identification. Eight of the designed SCAR markers were shown to be able to positively identify the corresponded cultivars as in RAPD.

This research focuses on the modeling of dendrimer molecules for their application as delivery vectors within drug and gene therapy systems. We examine how the architecture and composition of dendrimers affect their drug and gene binding efficacies along with their interactions with anionic bilayers. We specifically focus on how the weakly basic nature of dendrimer monomers and the addition of neutral grafts to dendrimer surface groups affect their interactions with drugs, linear polyelectrolytes, and bilayers. By using polymer self-consistent field theory (SCFT) to model such systems, we develop a computationally efficient means to provide physical insights into these systems, which are intended to guide dendrimer design for delivery applications.We study the conformational properties of weakly basic (annealed) polyelectrolyte dendrimers by developing a SCFT model that explicitly accounts for the acid-base equilibrium reaction of the weakly basic monomers. We specifically focus on the role of local counterion concentration upon the charge and conformations of the annealed polyelectrolyte dendrimers. We compare our results to existing polymer scaling theories and develop a strong stretching theory for the dendrimer molecules.We extend the previous study to model the interactions between weakly basic dendrimers and weakly acidic, hydrophobic drug molecules. We specifically examine the effects of excluded volume, electrostatic, and enthalpic interactions on the binding efficacies between dendrimers and drugs under a variety of dendrimer generations, solution pOH conditions, drug sizes, and Bjerrum length values.We study the role of neutral dendrimer grafts on the conformations and drug binding efficacies of dendrimers. We then elucidate how the observed conformational changes affect the charge of the dendrimers. Furthermore, we examine how the presence of grafts affects the steric, electrostatic, and hydrophobic interactions between the drugs and dendrimers under a variety of solution conditions. We compare our results with the binding efficacies observed for non-grafted dendrimers to delineate the conditions under which the grafted dendrimers are better suited as drug hosts.We include semi-flexible, anionic linear polyelectrolyte (LPE) molecules in our grafted dendrimer SCFT framework to model the interactions between dendrimers and negatively charged genetic materials. Specifically, we examine how neutral dendrimer grafts, LPE stiffness, and solution pOH affect the interactions between dendrimers and LPEs. We then use our SCFT potential fields as input into Monte Carlo simulations in order to determine the dendrimer-LPE potentials of mean force and the resulting loop and tail statistics of the dendrimer-adsorbed LPE chains.We incorporate a negatively charged bilayer into our grafted dendrimer SCFT framework to model dendrimer interactions with a cellular membrane. We specifically examine the role of dendrimer grafting length, solution pH, and membrane tension on such interactions. By comparing our results with SCFT calculations of fixed dendrimer conformations and hard sphere nanoparticles in the presence of membranes, we delineate the role of dendrimer flexibility and porosity on the interactions between dendrimers and anionic bilayers.
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碩士
國立中興大學
園藝學系所
96
The genetic diversity of 31 butterhead (BUT) and 26 crisphead (CRP) accessions were evaluated by morphological traits, RAPD (Random Amplified Polymorphic DNA) and SCAR (Sequence Characterized Amplified Regions). Data for 19 morphological traits were subjected to a genetic diversity analysis, after which a UPGMA cluster analysis was performed. The 57 accessions were clustered into 2 groups according to type with genetic similarity of 0.38. The first group included 15 accessions form Tainan and 3 accessions form the National Germplasm of the USA, which were clustered into 5 subgroups with the similarity beyond 0.61. The second group included 10 crisphead and 26 butterhead accessions, which were clustered into 6 subgroups with the similarity beyond 0.70. A total of 271 RAPD bands, with a mean band of 9.3 for each primer, were generated using 29 out of 130 primers in the RAPD analysis. The polymorphism was 53.9%. All accessions were clustered into 3 groups in the RAPD analysis. The first group included 13 crisphead accessions form the Taiwan agricultural research institute(TARI) with the similarity 0.86. The second group including 15 accessions form the Tainan district agricultural research and extention station(TDARES)、3 accessions form USA and 3 butterhead accessions with the similarity 0.88. The first, second and fourth subgroups included crisphead accessions. The third and fifth subgroups included 3 butterhead accessions. The third group was composed of 23 butterhead accessions with a the similarity of 0.82. In analyzing results from SCAR of butterhead and crisphead accessions, a total of 100 bands were generated using 21 out of 51 primer pairs. The means of bands generated by a primer pairs was 4.8 and the polymorphism was 73%. The 57 accessions were clustered into 3 groups. The first group included 13 crisphead accessions form the TARI with the similarity 0.76. The second group including 15 accessions form TDARES with a similarity of 0.88. The third group was composed of 26 butterhead and 3 accessions the USA with a similarity of 0.88. The analysis of RAPD and SCAR result in the main group of the dendrogram showed a similarity of 0.78~0.86 based on RAPD markers and which the SCAR markers displayed a similarity 0.68~0.78. The analysis of RAPD and SCAR result in the subgroup of the dendrogram showed a similarity of 0.84~0.96 based on RAPD markers and which the SCAR markers displayed a similarity 0.88~0.97. Lettuce germplasm could be identified by morphological characteristics such as leaf color, leaf texture and head type. The molecular markers could be used to cluster accessions into groups by SCAR and subgroups by RAPD successfully.