Bioluminescent calcium indicator protein and gold nanoparticle-enhanced bioluminescence

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WANG-THESIS-2018.pdf (2.18 MB)
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Wang, Zuhui
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http://hdl.handle.net/1853/59939
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Bioluminescent proteins have been extensively used as a light emission source for many fusion proteins and have a wide range of applications in imaging and cell signaling studies. In this study, the smallest known luciferase from the marine copepod, Gaussia princeps, was used to engineer novel bioluminescent sensors and novel methods to enhance bioluminescence intensity. In the First part of the study, two bioluminescent calcium sensors were developed, which were composed of a variant of Gaussia luciferase (sbGLuc) and the fluorescent calcium indicator protein, GCaMP6s. The two designs allowed bioluminescence resonance energy transfer (BRET) and they exhibited excellent dynamic range. In the Second part of the study, several strategies to enhance bioluminescence by conjugating sbGLuc with gold nanoparticles were carefully examined. On average, 26% enhancement on bioluminescence was achieved. The new sensors together with the gold nanoparticle-enhanced bioluminescence should be useful for various studies in various fields such as neuroscience and cell biology.
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2018-04-25
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