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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃筱鈞(Hsiao-Chun Huang) | |
| dc.contributor.author | Hao-Chun Fan | en |
| dc.contributor.author | 范浩俊 | zh_TW |
| dc.date.accessioned | 2022-11-23T08:58:09Z | - |
| dc.date.available | 2021-11-04 | |
| dc.date.available | 2022-11-23T08:58:09Z | - |
| dc.date.copyright | 2021-11-04 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-29 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79324 | - |
| dc.description.abstract | 不對稱細胞分裂是細胞分化的重要過程。借助合成生物學的概念，我們可以通過在對稱分裂的模式生物，如大腸桿菌，中實施核心的不對稱分裂調控網絡來重現該過程。先前，我們的研究中發現可以用新月柄桿菌中構築極性網絡的核心蛋白(PopZ)來建構最小的不對稱細胞分裂模型。PopZ會穩健的在大腸桿菌細胞中行成單一的極化點，並能在開啟下游的基因表達後藉由一個不會擴散的報導蛋白形成可視化的不對稱分模型，完成最小系統的建構。然而這個最小的不對稱性分裂系統對報導蛋白的表達水平波動很敏感，因此在這項研究中我們的目標是添加對向極的新系統來建立一個雙極的模型，以期建立一個更穩定的不對稱細胞分裂系統。此系統設計使用新月柄桿菌PodJ（另一種也在細胞極處自組織的支架蛋白）添加到先前的系統中，並利用一個PodJ 的結合蛋白(PleC)作為連接蛋白進行測試。本研究的結果表明，PodJ和PleC可以作為在大腸桿菌中重建的穩健不對稱細胞分裂的第二極。 | zh_TW |
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| dc.description.tableofcontents | 致謝 i 摘要 ii Abstract iii List of Abbreviations iv Contents v Figure list vii Table list ix 1. Introduction 1 2. Literature review 7 2.1 Synthetic biology 7 2.2 Asymmetric cell division 9 2.2.1 Introduction of asymmetric cell division 9 2.2.2 Mechanism of asymmetric cell distribution 11 2.2.3 Asymmetric cell division in C. crescentus 13 2.2.4 Asymmetric cell division in E. coli 18 2.3 C. crescentus PodJ 20 3. Method 21 3.1 In silico prediction 21 3.2 Molecular Cloning 22 3.2.1 Bacteria strain culture conditions 22 3.2.2 Plasmid 22 3.2.3 Plasmid mini preparation 23 3.2.4 Primer design 25 3.2.5 PCR 26 3.2.6 PCR clean up 27 3.2.7 Restriction enzyme digestion 28 3.2.8 Agarose gel electrophoresis 28 3.2.9 Gel purification 29 3.2.10 DNA ligation 30 3.2.11 Gibson assembly 30 3.2.12 BioBrick assembly 31 3.2.13 Promoter replacement 32 3.2.14 Chemical transformation 32 3.3 Mammalian cell line culturing 33 3.3.1 Hela cell culturing 33 3.3.2 Plasmid transfection 33 3.4 Fluorescent microscopy 34 3.4.1 Imaging 34 3.4.2 Analysis 34 4. Results and discussion 36 4.1 In silico prediction 36 4.1.2 C. crescentus PopZ 36 4.1.2 C. crescentus PodJ 37 4.1.3 C. crescentus PleC 37 4.2 The formation of second pole 38 4.2.1 Minimal self-aggregation domain of PodJ 38 4.2.2 PodJ without PopZ binding domain 41 4.3 The co-expression test of PodJ 43 4.3.1 Co-expression PodJ with PopZ 43 4.3.2 Co-expression PodJ with SpmX 44 4.3.3 Co-expression PodJ with PleC 45 5. Conclusion and future work 47 6. Figure 48 7. Reference 91 Appendix I Sequence of PodJ I Appendix II Sequence of PleC IV Appendix III Sequence of PopZ VII Appendix IV Python code VIII | |
| dc.language.iso | en | |
| dc.title | 在大腸桿菌中建立第二極以人工實現穩健不對稱細胞分裂 | zh_TW |
| dc.title | Establishment of the second pole for a robust synthetic asymmetric cell division in Escherichia coli | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳亘承(Hsin-Tsai Liu),史有伶(Chih-Yang Tseng) | |
| dc.subject.keyword | 不對稱細胞分裂,新月柄桿菌,細胞極化,大腸桿菌,最小系統, | zh_TW |
| dc.subject.keyword | asymmetric cell division,Caulobacter crescentus,cell polarization,Escherichia coli,minimal system, | en |
| dc.relation.page | 111 | |
| dc.identifier.doi | 10.6342/NTU202104321 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-10-30 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子與細胞生物學研究所 | |
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