| LDR | | 00000nam u2200205 4500 |
| 001 | | 000000433204 |
| 005 | | 20200225113726 |
| 008 | | 200131s2019 ||||||||||||||||| ||eng d |
| 020 | |
▼a 9781392300213 |
| 035 | |
▼a (MiAaPQ)AAI13886836 |
| 035 | |
▼a (MiAaPQ)ucsf:11796 |
| 040 | |
▼a MiAaPQ
▼c MiAaPQ
▼d 247004 |
| 082 | 0 |
▼a 574 |
| 100 | 1 |
▼a Feng, Siyu. |
| 245 | 10 |
▼a Split Fluorescent Protein Engineering and Applications. |
| 260 | |
▼a [S.l.]:
▼b University of California, San Francisco.,
▼c 2019. |
| 260 | 1 |
▼a Ann Arbor:
▼b ProQuest Dissertations & Theses,
▼c 2019. |
| 300 | |
▼a 126 p. |
| 500 | |
▼a Source: Dissertations Abstracts International, Volume: 80-12, Section: B. |
| 500 | |
▼a Publisher info.: Dissertation/Thesis. |
| 500 | |
▼a Advisor: Huang, Bo. |
| 502 | 1 |
▼a Thesis (Ph.D.)--University of California, San Francisco, 2019. |
| 506 | |
▼a This item must not be sold to any third party vendors. |
| 520 | |
▼a 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. |
| 590 | |
▼a School code: 0034. |
| 650 | 4 |
▼a Bioengineering. |
| 650 | 4 |
▼a Biology. |
| 650 | 4 |
▼a Biochemistry. |
| 690 | |
▼a 0202 |
| 690 | |
▼a 0306 |
| 690 | |
▼a 0487 |
| 710 | 20 |
▼a University of California, San Francisco.
▼b Bioengineering. |
| 773 | 0 |
▼t Dissertations Abstracts International
▼g 80-12B. |
| 773 | |
▼t Dissertation Abstract International |
| 790 | |
▼a 0034 |
| 791 | |
▼a Ph.D. |
| 792 | |
▼a 2019 |
| 793 | |
▼a English |
| 856 | 40 |
▼u http://www.riss.kr/pdu/ddodLink.do?id=T15491533
▼n KERIS
▼z 이 자료의 원문은 한국교육학술정보원에서 제공합니다. |
| 980 | |
▼a 202002
▼f 2020 |
| 990 | |
▼a ***1816162 |
| 991 | |
▼a E-BOOK |