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論文中文名稱:檢測鏈黴菌內兩個ttrA同源基因在接合作用上的功能 [以論文名稱查詢館藏系統]
論文英文名稱:To examine the conjugal functions of two ttrA homologs in SCP1 and S. coelicolor chromosome [以論文名稱查詢館藏系統]
院校名稱:臺北科技大學
學院名稱:工程學院
系所名稱:生物科技研究所
畢業學年度:97
出版年度:98
中文姓名:沈姵妏
英文姓名:pen-wen-sen
研究生學號:95688021
學位類別:碩士
語文別:中文
口試日期:2009-07-30
論文頁數:49
指導教授中文名:黃志宏
口試委員中文名:陳文盛;陳月茸
中文關鍵詞:接合作用鏈黴菌線型質體線型染色體
英文關鍵詞:conjugationstreptomyceslinear plasmidlinear chromosome
論文中文摘要:鏈黴菌具有線狀的染色體與質體,其未端都有末端蛋白以共價鍵附著在DNA的5'端,而其中部分的線型質體能促進接合作用,驅動質體與染色體的傳遞。在鏈黴菌接合作用的相關研究中,位於S. lividans染色體及其線型質體SLP2上的ttrA,此基因是唯一證實與線型DNA分子的傳遞有關的基因,當線型DNA分子上的ttrA基因被剔除後,會降低該DNA分子的接合作用的頻率。ttrA的同源序列出現在很多線型染色體及質體的末端區域,但在線型質體SCP1上卻是出現在中央的區域,且同源性較低。
本論文即是進一步檢測連黴菌不同的ttrA同源基因,藉由基因剔除的方式,去檢測這些突變株的接合作用能力是否會有所不同。在剔除S. coelicolor染色體上的ttrA部分,帶有插入SCP1的3456上,將染色體上ttrA刪除,與S. coelicolor M130交配後,染色體之間重組互換的頻率沒有明顯的改變,但如果是與S. lividans TK54交配,則其頻率降低約兩千倍;但刪除M145的ttrA後,不管是以SLP2或是SCP1質體驅動接合作用,和S. lividans TK54交配,或是當接受株和S. lividans 3200交配,其染色體之間重組互換的頻率都降低不到十五倍。在SCP1的部份,SCP1.136的剔除,會使得SCP1傳遞到recipient的能力降低二百倍,但是染色體之間重組互換的頻
率並沒有明顯的差異,這與Huang等人(2003)對SLP2剔除ttrA的接合作用結果相符;若是與染色體上ttrA缺失的突變株交配,則SCP1傳遞到接受株的能力會下降高達萬倍,顯示ttrA在接受株內對接合作用也是有影響的。
論文英文摘要:Streptomyces has linear chromosomes and linear plasmids, and terminal proteins are covalently linked at 5’ ends of these DNA. Part of Streptomyces linear plasmids own conjugal ability, they can transfer themselves and chromosomes into recipients. One of related gene of conjugal transfer of Streptomyces linear replicons is ttrA, and previously study showed that ttrA of S. lividans chromosome and SLP2 linear plasmid worked in cis for DNA transfer during conjugation. ttrA homologs are commonly located in the end of linear replicons of Streptomyces, but the linear plasmid SCP1 carries two ttrA homologous, with lower identity, located on the central region.
This study was analyzed the function of ttrA homologs of Streptomyces in conjugation by gene replacement. The ttrA of S. coelicolor chromosome showed the different effects on two strains, M145 without plasmids and 3456 with integrated SCP1. M145-
論文目次:中文摘要 i
英文摘要 ii
誌謝 iii
目錄 iv
表目錄 v
圖目錄 vi
第一章 緒論 1
第二章 材料與方法 11
第三章 實驗結果 11
第四章 討論 11
附錄
Appendix A. Media and buffer………………………………………………...11
Appendix B. E.Coli Competent cell preparation and transformation………….11 Appendix C. Plasmid isolation from E. coli……………………………………11
Appendix D. Preparation of Streptomyces protoplast and transformation……..11
Appendix E. Isolation the total DNA of Streptomyces………………………….11
Appendix F. Polymerase Chain Reaction……………………………………….11
Appendix G. Pulsed-field gel electrophoresis…………………………………..11
Appendix H. Southern hybridization……………………………………………11
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