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研究生:楊琳
研究生(外文):Lin Yang
論文名稱:轉染方法的探討與回顧
論文名稱(外文):Discussion and review of transfection methods
指導教授:侯劭毅侯劭毅引用關係
口試委員:黃志宏王勝仕
口試日期:2013-04-26
學位類別:碩士
校院名稱:國立臺北科技大學
系所名稱:生化與生醫工程研究所
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:82
中文關鍵詞:轉染技術病毒載體非病毒載體化學方法物理方法
外文關鍵詞:transfectionviral vector methodnon-viral vector methodchemical methodphysical method
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基因轉殖也可稱為轉染技術(transfection)是將目標基因引入到真核細胞,並使之增殖和表達的一種技術。核酸導入到細胞中是生命科學及基因工程中經常使用並且是最有價值的工具。基因轉殖除了可提供研究特定基因的功能,也有應用在農業和醫學上。基因轉殖已經利用病毒、非病毒載體的方法,以及物理方法。病毒載體是採用複製缺陷型病毒載體介導基因轉移,此類載體如腺病毒,腺相關病毒,逆轉錄病毒和單純皰疹病毒等病毒。病毒載體的高效率基因運送是他們主要的優點。非病毒載體包括陽離子脂質體,陽離子聚合物,合成肽和天然存在的化合物。這些非病毒載體高效率地運送基因到體外培養的細胞中。物理方法則是利用機械壓力、電擊或水動力使細胞膜瞬時地滲透,此時DNA傳送導入靶細胞。局部基因傳遞它們比病毒和非病毒為基礎的系統是簡單的和非常有效的。每種方法的轉染效率都不相同,同一種方法應用在不同研究上其轉染效率是不盡相同的,為了讓研究者可以對轉染技術所做的研究其轉染效率有所了解,對於設計實驗上有幫助,因此在本篇論文中,將提及的內容為: (i) 將介紹22種轉染方法; (ii) 解釋設計病毒,非病毒,物理基因傳遞方法的基本原理; (iii) 研究綜述轉基因技術的最新進展; (iv) 討論每一個最常用的基因傳遞方法的優點和缺點; 及(v) 提供未來前景。

Transfection is a technology of introducing the target gene into eukaryotic cell and the target gene amplified and expressed. Nucleic acids are transducted into cells, which is commonly used and the valuable tools in life science and genetic engineering. Gene transfection can not only be provided the research of specific gene function but also used in agriculture and medical science. Gene transfection has be search for using viral, non-viral vector and physical methods. Viral vector-mediated gene transfer employs replication-deficient viruses such as adenovirus, adeno-associated virus, retrovirus, herpes simplex virus, etc. High gene delivery efficiency of viral vectors of is their major advantage. Up to date, the nonviral vectors has been developed include cationic liposomes, cationic polymers, synthetic peptides and natural compounds. These nonviral vectors appear to be highly effective in gene delivery to cultured cells in vitro but are significantly less effective in vivo. Physical methods that are using physical force are mechanical pressure, electric shock or hydrodynamic force to transiently permeate the cell membrane to transfer DNA into target cells. They are simpler than viral- and nonviral-based systems and highly effective for localized gene delivery. The transfection efficiency of each methods are not the same. The same method applied in different studies that the results of the transfection efficiency are not the same. In order to make researchers better understanding the efficiency of the study which is made by transfection technology. For designing experiments it can be helpful. Therefore, the objectives in this review are (i) introducing twenty two transfection methods; (ii) explaining the rationale for the design of viral, nonviral and physical methods for gene delivery;(iii) providing a summary on recent advances in gene transfer technology; (iv) discussing advantages and disadvantages of each of the most commonly used gene delivery methods; and (v) providing future perspectives.

中文摘要 i
英文摘要 ii
誌謝 iv
目錄 v
表目錄 vii
圖目錄 viii
第一章 緒論 1
1.1基因轉殖(Transfection) 1
1.2基因轉殖技術之應用 2
第二章 基因重組病毒的方法 4
2.1腺病毒(Adenoviruses) 8
2.2腺相關病毒(Adenoassociated viruses) 11
2.3逆轉錄病毒(Retroviruses) 13
2.4慢病毒 (Lentiviruses) 16
2.5泡沫病毒載體 (Foamy Viral Vectors) 18
2.6單純皰疹病毒(Herpes simplex viruses) 和
牛痘病毒( Vaccinia viruses) 18
2.7猿猴病毒40 (Simian virus 40, SV40) 20
2.8 pED 20
2.9靶向基因轉移和可調控表達(Targeted gene transfer and
regulatable expression) 21
2.10不同的病毒系統整理與比較 23
第三章 化學的方法 25
3.1二乙基氨基乙基-葡聚醣 (DEAE (Diethylaminoethyl)
-dextran) 25
3.2磷酸鈣(Calcium-Phosphate) 26
3.3脂質體轉染(Lipofection) 27
3.4聚合物(Polymer) 30
第四章 物理的方法 33
4.1顯微注射(Microinjection) 33
4.2基因槍微粒輸送(Biolistic Particle Delivery) 33
4.3電穿孔(Electroporation) 35
4.4 Sonoporation 37
4.5激光輔助方法(Laser-Assisted Method) (Photoporation) 38
4.6磁轉 (Magnetofection) 39
4.7流體動力基因傳遞 (Hydrodynamic Gene Delivery)
(Hydroporation) 40
4.8細胞核基因殖入系統 (NucleofectorTM) 42
4.9針頭注射(Injection) 43
第五章 轉染方法的優缺點 49
第六章 結論及展望 53
參考文獻 56


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