No description available.

The purpose of this research was to study the hybridization of molecular beacons under different conditions and designs. Data collected suggest that the inconsistency found in the emission intensity of several of these probes may be caused by 3 important factors: length of the probe, nucleotide sequence and, the formation of an alternative complex structure such as a dimer. Of all three factors, dimer formation is the most troublesome, since it reduces the emission of the reporter molecules. A new probe design was used to reduce dimer formation. The emission signal of the improved probe was several folds stronger than those probes with the early design. In this research, dimer formation is detected, furthermore a new probe with a different design was tested. If dimer formation can be reduced molecular beacons can be integrated into more complex hybridization systems providing an important tool in research and diagnosis of genetic disorders.

The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18848号 / 医博第3959号 / 新制||医||1007(附属図書館) / 31799 / 京都大学大学院医学研究科医学専攻 / (主査)教授 岩田 想, 教授 岩井 一宏, 教授 楠見 明弘 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing

No description available.

Yes / FRET forms the basis for energy transfer in biological systems and organisms and it has become an investigative tool in the analysis of protein‐protein interactions and in the study of semiconductors (SC). Until now, FRET has been restricted to the simultaneous presence of both components in the same phase. Here, we report on the first successful prototype demonstrating interfacial FRET. This innovative FRET between inorganic SC‐nanoparticles and illuminating protein chimeras takes place across an oil/water interface. As a ′proof of concept′ oil droplets were stabilized by hydrophobin‐derivatives in aqueous solution. These proteins possess the ability to attach fused functional domains close to an interface. Moreover, an optically active nanostructure directly docks to the hydrophobin at the oil/water interface. Due to its modular design, this signal amplification array has the potential to be exploited in numerous fields ranging from biosensors, biotechnology to medical applications.

Yes / Dye-functionalized polymers have been extensively studied to understand polymer chain dynamics, intra or inter-molecular association and conformational changes as well as in practical applications such as signal amplification in diagnostic tests and light-harvesting antennas. In this work, the Förster resonance energy transfer (FRET) of dye-functionalized poly(2-ethyl-2-oxazoline) (PEtOx) was studied to evaluate the effect of dye positioning and polymer chain length on the FRET efficiency. Therefore, both α (initiating terminus)- or ω (terminal chain end)-fluorophore single labeled and dual α,ω-fluorescent dye labeled PEtOx were prepared via cationic ring opening polymerization (CROP) using 1-(bromomethyl)pyrene as the initiator and/or 1-pyrenebutyric acid or coumarin 343 as the terminating agent, yielding well-defined PEtOx with high labeling efficiency (over 91%). Fluorescence studies revealed that intramolecular FRET is most efficient for heterotelechelic PEtOx containing both pyrene and coumarin 343 fluorophores as chain ends, as expected. A strong dependence of the energy transfer on the chain length was found for these dual labeled polymers. The polymers were tested in both dilute organic (chloroform) and aqueous media revealing a higher FRET efficiency in water due to the enhanced emissive properties of pyrene. The application of dual labeled polymers as fluorescent probes for temperature sensing was demonstrated based on the lower critical solution temperature behavior of the PEtOx. Furthermore, these polymers could be successfully processed into fibers and thin films. Importantly, the fluorescence properties were retained in the solid state without decreasing the FRET efficiency, thus opening future possibilities for application of these materials in solar cells and/or sensors.

No description available.