Transcriptome analysis based on deep expressed sequence tag (EST) sequencing allows quantitative comparisons of gene expression across multiple species. Using pyrosequencing, we generated over 7 million ESTs from four stages of developing seeds of Ricinus communis, Brassica napus, Euonymus alatus and Tropaeolum majus, which differ in their storage tissue for oil, their ability to photosynthesize and in the structure and content of their triacylglycerols (TAG). The larger number of ESTs in these 16 datasets provided reliable estimates of the expression of acyltransferases and other enzymes expressed at low levels. Analysis of EST levels from these oilseeds revealed both conserved and distinct species-specific expression patterns for genes involved in the synthesis of glycerolipids and their precursors. Independent of the species and tissue type, ESTs for core fatty acid synthesis enzymes maintained a conserved stoichiometry and a strong correlation in temporal profiles throughout seed development. However, ESTs associated with non-plastid enzymes of oil biosynthesis displayed dissimilar temporal patterns indicative of different regulation. The EST levels for several genes potentially involved in accumulation of unusual TAG structures were distinct. Comparison of expression of members from multi-gene families allowed the identification of specific isoforms with conserved function in oil biosynthesis. In all four oilseeds, ESTs for Rubisco were present, suggesting its possible role in carbon metabolism, irrespective of light availability. Together, these data provide a resource for use in comparative and functional genomics of diverse oilseeds. Expression data for more than 350 genes encoding enzymes and proteins involved in lipid metabolism are available at the 'ARALIP' website ().

This research was designed to determine the conditions necessary to remove c-myc and 6x-His tags from a flavonol specific glucosyltransferase found in grapefruit (CP3GT) using thrombin in preparation for crystallization. X-ray crystallography of CP3GT crystals may elucidate structural features that account for flavonol specificity in some glucosyltransferase enzymes. A thrombin cleavage site was inserted into WT CP3GT and one mutant. Recombinant CP3GT was expressed in yeast and purified. Optimal conditions for thrombin digestion were explored. Digestion with 100U of thrombin for 2 hours at 4o C was optimal for removing tags from CP3GT. Storage at 4o C for 2 hours resulted in approximately 70% retention of activity. The effect of thrombin treatment on CP3GT activity was tested. Purified CP3GT protein with and without tags was tested for activity with the flavonol quercetin. Data showed no significant difference in overall activity between tagged and native protein.

京都大学 / 0048 / 新制・課程博士 / 博士(人間・環境学) / 甲第19797号 / 人博第768号 / 新制||人||185(附属図書館) / 27||人博||768(吉田南総合図書館) / 32833 / 京都大学大学院人間・環境学研究科共生人間学専攻 / (主査)教授 齋藤 治之, 教授 壇辻 正剛, 准教授 谷口 一美 / 学位規則第4条第1項該当 / Doctor of Human and Environmental Studies / Kyoto University / DGAM

Projektet handlar om utveckling av ett mätpixelbibliotek och en besöksräknare för företaget Askås. Detta är ett viktigt område på grund av att spårning av användardata på webben blir allt vanligare och låter oss förstå användarbeteende noggrannare. Det största problemet gällande projektet är att användarnas integritet måste skyddas samtidigt som insamlingen sker, därför behöver användarna ge sitt samtycke för insamling av data. Data som kategoriseras som personuppgifter krypteras och tas bort när de inte längre behövs. Syftet med detta examensarbete är att utveckla ett system som både är enkelt att använda och effektivt. För att lösa dessa problem implementeras JavaScript-kod på webbsidor som samlar in data. En server används för att ta emot data, bearbeta den och skicka till en databas. Systemets alla delar tillsammans kallar vi ett mätpixelbibliotek. Resultaten visar att systemet kan samla in och lagra användardata för vidare analys som till exempel för att räkna ut antalet besökare. Systemet testades på en webbsida och visade sig vara funktionellt. Analyser av den insamlade informationen kan ge insikter kring användarbeteende, vilket gör det möjligt att optimera webbplatsen. Sammantaget har projektet uppnått sina mål bestående av mätpixelbibliotek och besöksräknare. Trots det finns det plats för förbättringar och utveckling. / The project involves the development of a data collection system and a visitor-counter for the company Askås. This is an important area due to the tracking of user data on the web is becoming more common and it also allows us to understand user behavior more accurately. The biggest problem regarding the project is that when collecting users data we also have to protect their privacy, therefore the users need to give their consent for the collection of data. Data categorized as personal data is encrypted and deleted when no longer needed. The purpose of this thesis is to develop a system that is both useful and efficient. To solve these problems, JavaScript code that collects the data is implemented on web pages. A server is used to receive the data, process it and send it to a database. The parts of the system together is the data collecting system. The results show that the system can collect and store user data for further analysis like counting a websites number of visitors. The system was tested on a web page and was found to be functional. The analyzes of the collected information can provide insights into user behavior, which makes it possible to optimize the website. Overall, the project has achieved its goals, although there is room for improvement and development.

No / Sewer and gully flooding have become major causes of pollution particularly in the residential areas majorly caused by blockages in the water system and drainages. An effective way of avoiding this problem will be by deploying some mechanism to monitor gully pot water level at each point in time and escalating unusual liquid levels to the relevant authorities for prompt action to avoid a flooding occurrence. This paper presents a low cost power efficient gully pot liquid level monitoring technique. Passive RFID tags are deployed and signal variation from the Alien Reader Software are used to effectively estimate the level of liquid in the gully pot. The experimental set up is presented and an expository presentation is made of the passive tag design, modelled and simulated and adopted for same application.

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Home automation systems is something that has risen in popularity over the last decade. More and more real estate is equipped with various IoT systems in order to keep effective- ness and efficiency on a high level. With these systems comes highly advanced technical systems connected to various devices that report data collected from their environments. The data that is collected is then reformatted into tag lists in the software with a very specific name that follows a certain naming convention. The problem is that there is no transparency amongst companies when it comes to the standards they follow when it comes to naming their tags. This is investigated in this thesis by conducting a literature review and creating a design to interpret existing tag lists to make it easier to understand the structure. The thesis is made with the intent of answering the question to what the current state of research when it comes to standardization in tag lists. It is also exploring what a potential effective design for translating tags into a manageable format would look like. At the time of making this thesis there were not very extensive research when it comes to standards and naming convention in IoT tag lists, therefore a lot of the compiled research in this thesis refers to related ways of structuring names in other topics. The design was developed with the help of an scale-up company called Sensative that works with IoT tags on a daily basis. The end result of the design is a translation service that is able to sugges- tion relatable tags to translate when a user is manually renaming tags that are imported into the service. The design is built with the potential to later add artificial intelligence to suggest related tags for translation.

Escherichia coli survives in both nutrient rich nutrient-limited environments. As such, understanding the gene and protein level activity that occurs during stationary phase is considered an important aspect of bacterial survival. Escherichia coli has been studied for decades providing substantial insight into gene expression profiles in exponential phase and recently, during adaptation to stationary phase. This led to the discovery of RpoS as a growth phase-dependent sigma factor. Further studies indicated that there are many genes that are expressed in an RpoS-independent but stationary phase-specific manner. However, proteins represent the functional molecules of the cell. Additionally, protein expression does not always correlate with the corresponding gene expression patterns. Therefore, to obtain an in depth understanding of the proteins that play a role in long-term growth in E. coli, TMT- (Tandem Mass Tags) based quantitative proteomic analysis was performed to identify proteins that are preferentially expressed during prolonged starvation. We identified proteins that were both positively and negatively regulated by RpoS during stationary phase, such as GadA and TnaA, respectively. RpoS levels peaked during early stationary phase and declined thereafter. However, proteins that were RpoS-dependent continued to increase during prolonged stationary phase. Additionally, we identified proteins that were expressed in an RpoS-independent manner during stationary phase. This suggests that protein expression during early stationary phase is distinct from prolonged stationary phase. Furthermore, RpoS-independent proteins may also play an important role during long-term growth. / Thesis / Master of Science (MSc) / Escherichia coli adapts to shifts in nutrient availability using the alternative sigma factor RpoS which controls morphological and physiological changes. Although gene expression during growth has been extensively studied, comparable information regarding changes in protein abundance during prolonged incubation is not available. We employed a quantitative proteomics approach to identify proteins that are preferentially expressed during stationary phase in E. coli. We identified classes of proteins that are upregulated and downregulated by RpoS in addition to proteins regulated independently of RpoS. Global analysis of protein expression during growth can aid in understanding the adaptation of E. coli under starvation conditions.