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<p>Totally 84 differentially expressed mRNA clones from infective L3 larvae of the parasite <em>Haemonchus contortus</em>, a blood sucking nematode,<em> </em>were analyzed with single strand hybridization assay (SSH). Altogether 79 clones were sequenced, edited, and compared with proteins found via BLAST in GeneBank. The aim was to investigate gene expression and potential protein expression following storage at 5 °C for 32 weeks. mRNA was extracted from fresh and stored L3. The SSH derived products were cloned into <em>E. coli,</em> purified and sequenced with Big Dye Terminator v3.1, and then compared with uploaded sequences in GeneBank. BLAST showed 59 (70 %) reliable protein results, where 39 (66 %) were of nematode origin.. Three sequences (4 %) were recognized as <em>H. contortus</em>-related metabolic proteins. Further research is necessary to elucidate the role of these proteins in relation to storage<em>. </em>Many of the sequences (36 %) are also present in other nematodes, such as <em>Caenorhabditis briggsae </em>and <em>C. elegans,</em> where whole-genome projects have been conducted. Bigger and more accurate databases need to be developed. Maybe the most significant future project is to sequence the whole genome of <em>H. contortus</em>.</p>

Vénus, deuxième planète du système solaire, souvent appelée sœur de la Terre, car présentant des caractéristiques de taille et de masse similaires, est fort différente de notre planète. En effet, son atmosphère est beaucoup plus dense et dynamiquement active que celle de la Terre. Dans ce travail, nous nous concentrerons sur la région s'étendant de 70 km à 180~km d'altitude : la mésosphère et la basse thermosphère. SOIR, acronyme de Solar Occultation in the InfraRed, est un instrument russo-franco-belge embarqué à bord de la mission Venus Express de l'Agence Spatiale Européenne. Il utilise un réseau à échelle comme élément diffractant. La plage de nombre d'onde mesurable par SOIR s'étend de 2200 cm-1 à 4400 cm-1, ou 4.3 µm à 2.2 µm en longueur d'onde. Les ordres de diffraction utiles varient de 101 à 194. Un filtre acousto-optique - AOTF - est utilisé comme passe-bande pour sélectionner les ordres de diffraction. La résolution de l'instrument varie de 0.13 à 0.24 cm-1. Du fait d'un temps de développement raccourci, presque toutes les calibrations de l'instrument ont dû être réalisées en vol. Elles comprennent les calibrations relatives au réseau à échelle (angle exact de Blaze et fonction de Blaze), au détecteur (non-uniformité pixel à pixel, courbe de sensibilité spectrale, relation entre les pixels et les nombres d'onde, résolution de l'instrument, intervalle d'échantillonnage spectral, rapport signal sur bruit, background thermique) et à l'AOTF (relation entre la radio-fréquence d'excitation de l'AOTF et les nombres d'onde, fonction de transfert de l'AOTF). L'approche et la réalisation de ces différentes calibrations sont présentées dans le présent travail. Les caractéristiques de l'instrument y sont également décrites. Un algorithme d'inversion spectrale a été développé pour le cas spécifique de SOIR. Tenant compte du mode de mesure, l'occultation solaire, la méthode de la pelure d'oignons a été implémentée dans un algorithme dit de l'Estimation Optimale. Cette méthode permet d'inverser l'ensemble des mesures spectrales en une fois, et également d'en tirer d'autres informations, comme l'amélioration de certaines caractéristiques instrumentales. Les paramètres qui sont ajustés au sein de l'algorithme sont la densité de la ou des espèces absorbant dans la région spectrale concernée, la température, les paramètres de la ligne de base, qui permettent de déterminer les caractéristiques des aérosols, le déplacement Doppler des raies qui trouve principalement son origine dans la vitesse de déplacement du satellite, et l'amélioration de certaines calibrations. Une étude de sensibilité des différents paramètres de l'algorithme est également présentée, ainsi qu'une évaluation des erreurs instrumentales systématiques. Dans l'ensemble des spectres enregistrés durant les 4 premières années de la mission, des profils verticaux de CO2, CO, H2O, HDO, SO2, H2SO4, HCl et HF ont été obtenus. Des valeurs limites de densité de OCS, H2CO, O3 et CH4 ont également été calculées. Les résultats concernant le dioxyde de carbone sont développés dans le texte. Des profils verticaux de CO2 s'étendant de 70 km à 180 km d'altitude sont analysés en profondeur. Ils sont comparés aux profils dits hydrostatiques, et des hypothèses quant à la dynamique agissant au niveau des deux terminateurs de Vénus sont formulées. / Venus, second planet of the solar system, is often seen as the sister planet of the Earth. In terms of size and mass, they are indeed very similar, but the Venus atmosphere is much thicker and active. The altitude region extending from 70 km to 180~km is studied in this thesis, namely the mesosphere and the lower thermosphere. SOIR, which stands for Solar Occultation in the InfraRed, is a Russian / French / Belgian instrument flying on board of the Venus Express (VEX) spacecraft of the European Space Agency. The diffracting device of SOIR is an echelle grating. The wavenumber region studied ranges from 2200 cm-1 to 4400 cm-1, or 4.3 µm to 2.2 µm in wavelength. The diffraction orders used with SOIR vary from 101 to 194. To select the required echelle grating diffraction order, an Acousto-Optical Tunable Filter - AOTF - is located after the entrance slit of the instrument. The instrument resolution varies from 0.13 to 0.24 cm-1. SOIR was developed in a very short time. Thus, virtually all the calibrations had to be made in-flight. These concern the echelle grating (exact Blaze angle computation, Blaze function), the detector (pixel-to-pixel non-uniformity, spectral sensitivity curve, pixel-to-wavenumber relationship, instrument resolution, spectral sampling interval, signal-to-noise ratio, thermal background) and the AOTF (wavenumber to AOTF frequency relation, AOTF transfer function). The procedure for and the computation of these calibrations are described in this work, as well as the instrument characteristics. A spectral inversion algorithm was developed specifically for the SOIR measurement technique: the solar occultation. The onion peeling method is implemented using the Optimal Estimation Method. It allows the inversion of the spectral data in one go, and also the enhancement of some instrumental characteristics. The algorithm variables are the densities of the species absorbing in the diffraction order, the temperature of the atmosphere under study, the spectral background parameters, that allow the determination of the Venus aerosols characteristics, the Doppler shift (mainly linked to the shift induced by the satellite displacement), and the improvement of some instrumental calibrations. A sensitivity study on the algorithm parameters is also presented, and the instrumental systematic errors are investigated. Vertical profiles of CO2, CO, H2O, HDO, SO2, H2SO4, HCl and HF are derived from the spectra measured during the first 4 mission years. Upper limits on OCS, H2CO, O3 and CH4 have also been calculated. We focus on the carbon dioxide results in the present study. A selection of vertical profiles extending from 70 km to 180 km are analyzed in details. They are compared to the hydrostatic profiles, and propositions concerning the terminators' dynamics are formulated.

Totally 84 differentially expressed mRNA clones from infective L3 larvae of the parasite Haemonchus contortus, a blood sucking nematode, were analyzed with single strand hybridization assay (SSH). Altogether 79 clones were sequenced, edited, and compared with proteins found via BLAST in GeneBank. The aim was to investigate gene expression and potential protein expression following storage at 5 °C for 32 weeks. mRNA was extracted from fresh and stored L3. The SSH derived products were cloned into E. coli, purified and sequenced with Big Dye Terminator v3.1, and then compared with uploaded sequences in GeneBank. BLAST showed 59 (70 %) reliable protein results, where 39 (66 %) were of nematode origin.. Three sequences (4 %) were recognized as H. contortus-related metabolic proteins. Further research is necessary to elucidate the role of these proteins in relation to storage. Many of the sequences (36 %) are also present in other nematodes, such as Caenorhabditis briggsae and C. elegans, where whole-genome projects have been conducted. Bigger and more accurate databases need to be developed. Maybe the most significant future project is to sequence the whole genome of H. contortus.

Metabolic engineering has enabled the advancement of biotechnology over the past few decades through the use of cells as biochemical factories. Cellular factories have now provided many new safe and sustainable routes to fuels, pharmaceuticals, polymers, and specialty chemicals. Many of these successes have been achieved using the yeast Saccharomyces cerevisiae, which has been used and shaped by humans for millennia for the production of food and drink. Consequently, S. cerevisiae is one of the most studied eukaryotic organisms in existence, and has established genetic tools for engineering efforts. However, despite the many achievements in metabolic engineering of S. cerevisiae, it is still significantly more difficult to engineer than its prokaryotic counterpart, Escherichia coli. As a result, there is an unmet need to further develop genetic tools in yeast and to do so in the context of metabolic pathway engineering. The work presented here addresses this need through the study and engineering of heterologous gene expression. First, a new biosynthetic pathway is engineered for the production of muconic acid in yeast. Muconic acid is a dicarboxylic acid that can serve as a platform chemical for the production of several bio-polymers. The final muconic acid producing strain was developed through significant metabolic engineering efforts and reached a titer of nearly 141 mg/L muconic acid, a value nearly 24-fold higher than the initial strain. Second, a new method of engineering promoters is presented that allows for increased expression of native promoters and the de novo design of synthetic promoters. The highest expression synthetic promoter is within the top 6th percentile of native yeast promoters. Third, a study of native and synthetic terminators for heterologous gene expression is completed for the first time. This study demonstrates that terminators can tune heterologous expression by as much as an order of magnitude. Fourth, a comparative study of plasmid components dissects the different contributions to plasmid burden, copy number, and gene expression level. Collectively, this work represents a significant step forward in the metabolic engineering of yeast through the establishment of a new pathway and the study and engineering of new tools for heterologous gene expression. / text

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第22707号 / 農博第2423号 / 新制||農||1080(附属図書館) / 学位論文||R2||N5300(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 本田 与一, 教授 田中 千尋, 教授 吉村 剛 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

Master of Science / Genetics Interdepartmental Program - Plant Pathology / Frank F. White / Sanzhen Liu / Plants and bacterial pathogens are in constant co-evolution to survive and sustain the next generation. Plants have two well-characterized levels of active defense -pathogens-associated molecular patterns (PAMPs)-triggered immunity (PTI) and effectors-triggered immunity (ETI). Some plants that are hosts for bacterial pathogens employing type three secretion system transcription activator-like (TAL) effectors have evolved a unique form of ETI, namely TAL effector-mediated ETI. TAL effectors induce expression of specific disease susceptibility (S) genes. Rice and pepper have evolved resistance genes termed terminator (T) genes, which have promoters that bind TAL effectors and, upon expression of the T gene, elicit a hypersensitive reaction (HR) and cell death. Only five T genes have been cloned, and the origin of most T genes is unknown. To determine the presence of candidate T genes in other plants species, a bioinformatics-based mining was designed. The basic approach utilized three structural features common to four terminator genes: a short trans-membrane domain, a secretion signal domain, and a length of <200 amino acid residues. Soybean was chosen as the test plant species, and 161 genes were retrieved that fulfilled the three parameters using R and Perl software programs. Further, functional annotation of candidate genes was conducted by comparisons to genes in public databases. Major classes of proteins found included unique and hypothetical, defense/stress/oxidative stress associated, DNA-binding, kinases, transferases, hydrolases, effector-related tRNA splicing, and F- box domain proteins. The potential T genes will serve as candidates for experimental validation and new resources for durable resistance strategies in crop species.

This research investigates partial discharge phenomena of on-site power transmission cables of 69 kV and 161 kV by inspecting the data measured under various environments. Effective procedures and methods are suggested for estimating insulation condition of power transmission cables. To tackle the problem of interference of the environmental noises to the on-site power transmission cables, a detection method is presented based on the relationship among the amplitude, phase, and focused area of electrical signals. The difference between the outer and inner electrical signals in power transmission cable is analyzed to improve the accuracy of pattern recognition. In addition, the insulation conditions of the cable joint and the cable terminator are evaluated by comparing results of electrical and non-electrical detections. Finally, a standard procedure with some key points of inspection processes under various environments is suggested to evaluate the cable insulation degradation based on the inspection data obtained from various environments.

Thesis (Ph. D.)--University of Washington, 1996. / Vita. Includes bibliographical references (leaves [115]-121).

Thesis (Ph.D.)--University of Alabama at Birmingham, 2007. / Title from PDF title page (viewed on Sept. 16, 2009). Includes bibliographical references.