Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 10 of 10 for “"Halotag"”.
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Turnover Rate of The Neuronal Connexin Cx36 In Hela Cells
… culture cells and whole organs. We utilized HaloTag technology to study the turnover rate of Cx36 in transiently transfected HeLa cells. The HaloTag protein forms irreversible covalent bonds with chloroalkane ligands, allowing pulse-chase experiments to be performed very specifically. The …
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Synthesis and characterisation of peptide-based probes for quantitative multicolour STORM imaging
… to activator and emitter dyes. Peptides with HaloTag ligands attached were labelled with various activator and emitter pairs and used to label HaloTag fusions of S. pombe and mouse embryonic stem cells. These peptides were found to bind non-specifically to various areas of both cell types, and …
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Two-Color Fluorescent Analysis of Connexin 36 Turnover and Trafficking – Relationship to Functional Plasticity
… communication. We utilized Cx36 with an internal HaloTag to study Cx36 turnover and trafficking in cultured cells to discriminate newly formed and pre-existing Cx36. New Cx36 in cargo vesicles was added directly to existing gap junctions and newly made Cx36 was not confined to points of addition, …
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Biophysical characterization of the Escherichia coli RNA degradosome
… RNA degradosome fluorescently labeled using a HaloTag showed discrete particles of the enzyme complex on a supported lipid bilayer on glass. To enable in situ studies of the degradosome, the HaloTag was encoded at the end of the rne gene within the E. coli chromosome using CRISPR/Cas. In order …
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Anomalous diffusion of mRNA in the cytoplasm of HeLa cells
… loops bound to coat proteins tagged with the HaloTag-JF646 fluorophore. We then used single-particle tracking to obtain trajectories of individual molecules. Trajectories were analyzed in terms of their mean-squared displacement (MSD) and power spectral density (PSD). We observed non-ergodic, …
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The role of adaptor protein Mal/TIRAP in transducing TLR signalling
… CRISPR/Cas9 HDR approaches to introduce a HaloTag onto the endogenous Tirap gene in immortalized bone marrow-derived macrophages (iBMMs). The introduction of the HaloTag did not affect the function of the Mal/TIRAP protein which retained its wild-type function (validated by LPS-induced …
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Understanding The Role of Arglu1 In Interferon Signaling Activation In Breast Cancer
… To identify RNAs directly bound by ARGLU1, Halotag-fused ARGLU1 was expressed in MCF7 cells for conducting CLIP-seq. These results elucidate unappreciated regulators of type I IFN signaling in breast cancer, offering potentially new targets to overcome cancer resistance to immune checkpoint …
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A neuron within a neuron: the effect of narrow axonal ER tubules on molecular movement
… On sparse chemical labelling of a luminal HaloTag protein, I found two classes of single-particle movement: slower and relatively bright particles that showed largely unidirectional movement, and faster less brightly labelled particles. The slower class showed no bias in the direction of …
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Novel Optical Tools to Investigate Presenilin Control of Neurotransmission
… the low affinity ER-GCaMPs in which we fused the HaloTag protein to allow for direct reporting of ER Ca<sup>2+</sup> levels. We coupled this new indicator with in-cell calibrations at physiological temperatures to create a highly robust readout of ER Ca<sup>2+</sup> across various cell types. Our …
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Investigating TLR-4 signalling in response to protein ligands
… transduced with a Mal/TIRAPHALO construct. The halotag was fluorescently labelled then the cells were stimulated with TLR-4 ligands, such as LPS, fixed at different time points, then imaged. Total internal reflection fluorescence (TIRF) microscopy was used to image the plasma membrane and …