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Split Fluorescent Protein Engineering and Applications
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| 자료유형 | 학위논문 |
|---|---|
| 서명/저자사항 | Split Fluorescent Protein Engineering and Applications. |
| 개인저자 | Feng, Siyu. |
| 단체저자명 | University of California, San Francisco. Bioengineering. |
| 발행사항 | [S.l.]: University of California, San Francisco., 2019. |
| 발행사항 | Ann Arbor: ProQuest Dissertations & Theses, 2019. |
| 형태사항 | 126 p. |
| 기본자료 저록 | Dissertations Abstracts International 80-12B. Dissertation Abstract International |
| ISBN | 9781392300213 |
| 학위논문주기 | Thesis (Ph.D.)--University of California, San Francisco, 2019. |
| 일반주기 |
Source: Dissertations Abstracts International, Volume: 80-12, Section: B.
Publisher info.: Dissertation/Thesis. Advisor: Huang, Bo. |
| 이용제한사항 | This item must not be sold to any third party vendors. |
| 요약 | Self-associating split fluorescent proteins (SAsFPs) have been widely used for labeling proteins, visualization of subcellular protein localization, scaffolding protein assembly and detecting cell-cell contacts. This PhD thesis research has been focused on expanding the toolbox of SAsFPs by adding more colors, increasing the brightness and empowering them with various biophysical properties. Utilizing a screening platform for the direct engineering of SAsFPs, we have generated a yellow-green split-mNeonGreen21-10/11, a 10-fold brighter red-colored split-sfCherry21-10/11 and demonstrated dual-color endogenous protein tagging using sfCherry211 and GFP11. However, the newly developed SAsFPs have suffered from sub-optimal fluorescence signal. By investigating the complementation process, we have employed two approaches to improve the overall brightness of SAsFPs: assistance through SpyTag/SpyCatcher covalent interaction and directed evolution for complementation-enhancing mutations. The latter strategy has then yielded two red-colored split sfCherry3 variants with substantially enhanced endogenous protein labeling performance. Based on sfCherry3, we have further developed a red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP) for multiplexed visualization of neuronal synapses in living animals, demonstrating the broad applications of SAsFPs. |
| 일반주제명 | Bioengineering. Biology. Biochemistry. |
| 언어 | 영어 |
| 바로가기 | 
: 이 자료의 원문은 한국교육학술정보원에서 제공합니다. |
