Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23135
DC FieldValueLanguage
dc.contributor顏宏真zh_TW
dc.contributor周映孜zh_TW
dc.contributor.advisor黃皓瑄zh_TW
dc.contributor.author傅碧汝zh_TW
dc.contributor.authorFu, Bi-Juen_US
dc.contributor.other中興大學zh_TW
dc.date2011zh_TW
dc.date.accessioned2014-06-06T07:19:37Z-
dc.date.available2014-06-06T07:19:37Z-
dc.identifierU0005-2610201014152400zh_TW
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dc.identifier.urihttp://hdl.handle.net/11455/23135-
dc.description.abstract農桿菌(Agrobacterium tumefaciens)為一種植物病原菌菌,屬於根瘤菌科農桿菌屬,革蘭氏陰性桿菌,菌體內含有一環狀染色體及一線狀染色體、二種質體(pTiC58與pAtC58)。Ti質體會使農桿菌感染植物後,經由Ti質體內的T-DNA的轉移,進而造成植物產生冠瘿狀腫瘤,也可做為產生轉基因作物的工具。目前推論農桿菌會藉由第四型分泌系統(type IV secretion system,T4SS)中的T線毛(T-pilus),將T-DNA與重要的致病毒蛋白(virulence protein)送出菌體外,進而進入植物細胞。已知T4SS是由VirB1-11及VirD4蛋白質所組成一穿膜的蛋白質複合體,其中T線毛主要由VirB2蛋白質及VirB5、VirB7蛋白質所組成。之前研究指出在酵母菌雙雜交系統(yeast two-hybrid)及in vitro中,阿拉伯芥中的RTNLB1 [BTI1 (VirB2-interacting protein 1)]、RTNLB2 (BTI2)及RTNLB4 (BTI3)這三個蛋白質,會兩兩互相結合,且可和VirB2蛋白質結合。本研究中,分別將RTNLB2蛋白質或在RTNLB1、RTNLB2或RTNLB4蛋白質的N端接上了T7標籤序列或His標籤序列大量表現於轉殖植物中,經農桿菌感染測試後得知皆可增加被農桿菌感染的效率,進一步確認RTNLB1、2、4蛋白質在農桿菌感染植物之過程中皆扮演相當重要之角色。並檢測RTNLB家族中的其他成員是否也有參與此過程,分別測試rtnlb3、rtnlb5、rtnlb6或rtnlb7突變株被農桿菌感染後之效率,結果顯示其產生腫瘤及短暫表現T-DNA的效率與野生株相似或比野生株些微增加或降低,故推測RTNLB3、RTNLB5、RTNLB6及RTNLB7基因在農桿菌感染植物之過程扮演類似之功能。另外,為了瞭解RTNLB基因家族在植物生長發育中可能扮演的角色與功能,本研究分析RTNLB基因家族的同源性關係;及利用Genevestigator軟體,得知RTNLB1-21基因在阿拉伯芥不同組織部位中的表現皆有不同。也利用逆轉錄聚合酶連鎖反應,得知RTNLB3、RTNLB5、RTNLB6及RTNLB7基因在阿拉伯芥不同組織部位均有表現。由以上結果可推測RTNLB基因在植物生長發育中相當重要,對其所具有生理意義值得更深入地研究探討。未來可以利用帶有標籤序列的RTNLB1、2、4轉殖株,進一步觀察在農桿菌感染植物時,RTNLB1、2、4蛋白質在細胞中分布之位置,或與之結合的植物蛋白質進行免疫沈澱並分離。以藉此更進一步釐清在農桿菌感染植物時,RTNLB1、2、4蛋白質所扮演的角色,與RTNLB蛋白質在植物中的功能。期許可以更廣泛的利用農桿菌來獲得轉殖物種,並提高轉殖的效率。zh_TW
dc.description.abstractAgrobacterium tumefaciens is a plant pathogen resided in the soil and is a Gram negative bacteria. The A. tumefaciens contains a circular and a linear chromosome, and two plasmids, pTiC58 and pAtC58. The A. tumefaciens infects plants and causes crown gall disease due to the transfer of the T-DNA (transferred DNA) on the Ti (tumor-inducing) plasmid. Because of the ability to transfer DNA between different kingdoms, the A. tumefaciens is also used to generate genetic modified crops. The A. tumefaciens utilize the type IV secretion system (T4SS) to secrete T-DNA and virulence proteins from the bacterium into plant cells. The T4SS contains a transmembrane protein complex composed of the VirB1-11 and VirD4 proteins and the T-pilus. The VirB2, VirB5, and VirB7 are the major and minor components of the T-pilus, respectively. A previous study has shown that three Arabidopsis plant proteins, RTNLB1 [BTI1 (VirB2-interacting protein 1)], RTNLB2 (BTI2), and RTNLB4 (BTI3), interacted with each other, and interacted with the VirB2 protein in the yeast two-hybrid system and in vitro. In order to understand the roles of RTNLB1, 2, and 4 during A. tumefaciens infection process, I generated overexpression RTNLB1, 2, or 4 Arabidopsis transgenic plants and performed A. tumefaciens infection assays. The overexpression RTNLB1, 2, or 4 transgenic plants showed higher transformation efficiencies than wild-type plants, suggesting RTNLB1, 2, and 4 all participate in A. tumefaciens infection process. Additionally, in order to test if other members of RTNLB are involved in A. tumefaciens transformation process, rtnlb3, rtnlb5, rtnlb6, or rtnlb7 T-DNA insertion mutants were obtained and tested further. Some of the tested rtnlb3, 5, 6, and 7 mutants showed similar transformation efficiencies as the wild-type, while others shower slightly increased or decreased transformation efficiencies compared to wild-type plants. These results suggest the RTNLB3, 5, 6, and 7 might be functional redundant in A. tumefaciens transformation process. In order to understand the possible functions of RTNLB genes in plant growth and development, the Genevestigator software was used to analyze the expression patterns and levels of RTNLB1-21 genes in different tissues of Arabidopsis plants. Results from various microarray experiments showed that RTNLB1-21 genes express differently in different tissues of Arabidopsis. Additionally, reverse transcription polymerase chain reaction (RT-PCR) results showed that RTNLB3, RTNLB5, RTNLB6 and RTNLB7 genes express ubiquitously in various plant tissues. In conclusion, results shown in this study suggest that RTNLB1-21 genes are important during plant growth and development. In the future, we could utilize immunoprecipitation approaches to understand further the functions of RTNLB1, 2, and 4 in Agrobacterium infection process and in plants.en_US
dc.description.tableofcontents中文摘要…………………………….………………………I Abstract…………………………...………………………II 目錄…………………………………….……………………III 表目錄……………………………………….………………IX 圖目錄………………………………………….……………X 附錄……………………………………………………….…XII 壹、前言…………………………………… ……………………… 1 一、農桿菌發展歷史與簡介……………………………………………1 二、農桿菌感染植物的過程……………………………………………2 1、農桿菌附著與辨認植物細胞……………………………………2 2、農桿菌中誘導表現vir基因……………………………………4 3、農桿菌中T-DNA的產生…………………………………………5 4、農桿菌中T-DNA與Vir蛋白質之轉移……………………………5 5、T-DNA送入植物細胞核…………………………………………8 6、T-DNA嵌入植物染色體中與表現………………………………9 三、RTNLB (reticulon-like)蛋白質的介紹…………………………10 1、RTNLB蛋白質的起源與種類……………………………………10 2、RTNLB蛋白質的結構……………………………………………11 3、RTNLB蛋白質在細胞中的位置與功能…………………………12 四、研究動機…………………………………………………………14 貳、材料與方法…………………………………………………………16 一、實驗材料……………………………………………………………16 1、植物……………………………………………………………16 2、菌種……………………………………………………………16 2.1、用於分析阿拉伯芥植物性狀之菌種…………………16 2.2、用以產生阿拉伯芥轉殖植物之菌種…………………16 3、培養基…………………………………………………………22 3.1、LB固態及液態培養基…………………………………22 3.2、523固態及液態培養基…………………………………23 3.3、B5固態培養基…………………………………………23 3.4、MS固態培養基…………………………………………23 3.5、CIM (callus induction medium)固態培養基………23 二、實驗方法:…………………………………………………………23 1、構築用於製造轉殖植物的菌體之製備方法…………………23 1.1、大腸桿菌勝任細胞(E. coli competent cell)之製備……24 1.2、大腸桿菌之熱休克轉型法……………………………24 1.3、農桿菌勝任細胞之製備………………………………24 1.4、農桿菌之電穿孔轉型法………………………………24 1.5、細菌內質體DNA之抽取法………………………………25 1.6、構築可於植物細胞中大量表現RTNLB2基因的質體…25 1.7、構築可於植物細胞中大量表現T7-tagged-RTNLB重組蛋白質之質體……26 1.8、可大量表現His-tagged- RTNLB重組蛋白質之質體構築……28 1.9、構築可於植物細胞中用RTNLB1啟動子區域表現GUS蛋白質之質體……29 1.10、限制酶之切割作用(restriction enzyme digestions) ……………………29 1.11、連接酶之連接反應(ligation reactions) ……………………………………29 1.12、利用菌落PCR篩選具有內插子(insert)DNA片段的細菌菌落……30 1.13、水平膠體之電泳分析(agarose gel electrophoresis) ……30 1.14、水平膠體內DNA片段及聚合酶連鎖反應(PCR)產物之純化與萃取…….30 2、植物材料之製備方法…………………………………………31 2.1、阿拉伯芥種子之消毒方法……………………………31 2.2、阿拉伯芥植物之培養方法……………………………31 2.3、阿拉伯芥植物之轉殖方法……………………………31 2.4、阿拉伯芥植物之篩選方法……………………………31 3、RTNLB轉殖株與T-DNA insertion突變株之分子檢測………32 3.1、T-DNA insertion突變株的基因組去氧核醣核酸(genomic DNA )之萃取……32 3.2、阿拉伯芥植物的核醣核酸(RNA)之萃取………………33 3.3、DNA或RNA濃度的定量…………………………………33 3.4、T-DNA insertion突變株的genomic DNA PCR………33 3.5、逆轉錄聚合酶連鎖反應(reverse transcription polymerase chain reaction,RT-PCR) ……………………………34 4、RTNLB轉殖株之蛋白質檢測……………………………………35 4.1、阿拉伯芥轉殖植物的蛋白質之萃取…………………35 4.2、阿拉伯芥轉殖植物的蛋白質之濃度測定……………36 4.3、阿拉伯芥轉殖植物的蛋白質之電泳分析(SDS-polyacrylamide gel [SDS-PAGE] analysis) ………………………36 4.4、西方轉漬法(Western blot analysis) ……………37 5、T-DNA insertion突變株、RTNLB轉殖株及ProRTNLB1::GUS轉殖株之性狀分析…38 5.1、腫瘤性狀分析(tumorigenesis assay) ……………38 5.2、短暫表現T-DNA性狀分析(transient transformation assays) ……38 5.3、觀察ProRTNLB1::GUS轉殖株在植物不同生長階段之GUS蛋白質表現情形……39 6、利用軟體分析阿拉伯芥中RTNLB基因家族……………………39 6.1、使用MEGA 4軟體分析RTNLB同源性關係………………39 6.2、利用Genevestigator軟體分析RTNLB16至RTNLB21基因於各個植物組織部位及不同環境處理之表現情形……………………39 參、結果………………………………………………………………40 一、RTNLB2、T7-tagged RTNLB2或His-tagged RTNLB2大量表現的轉殖植物產生種瘤及短暫表現T-DNA的效率皆增加……………………40 1、RTNLB2、T7-tagged RTNLB2或His-tagged RTNLB2大量表現的轉殖株之RTNLB2 RNA累積量的分析……………………………………40 2、RTNLB2、T7-tagged RTNLB2或His-tagged RTNLB2大量表現轉殖株的腫瘤及短暫表現T-DNA之性狀分析……………………………41 二、T7-tagged RTNLB1或His-tagged RTNLB1大量表現的轉殖植物產生種瘤及短暫表現T-DNA的效率皆增加………………………………42 1、 T7-tagged RTNLB1大量表現或His-tagged RTNLB1大量表現的轉殖株之RTNLB1 RNA累積量分析……………………………42 2、 T7-tagged RTNLB1大量表現轉殖株或His-tagged RTNLB1大量表現轉殖株的腫瘤及短暫表現T-DNA之性狀分析…………43 三、T7-tagged RTNLB4大量表現的轉殖植物產生種瘤及短暫表現 T-DNA 的效率皆增加………………………………44 1、T7-tagged RTNLB4大量表現的轉殖株之RTNLB4 RNA累積量分析………44 2、T7-tagged RTNLB4大量表現轉殖株的腫瘤及短暫表現T-DNA之性狀分析……45 四、rtnlb3、rtnlb5、rtnlb6及rtnlb7的T-DNA insertion突變株被農桿菌感染後產生腫瘤及短暫表現T-DNA的性狀分析…………45 1、rtnlb3突變株被農桿菌感染後,產生腫瘤及短暫表現T-DNA的效率無顯著增加……46 2、rtnlb5突變株被農桿菌感染後,產生腫瘤及短暫表現T-DNA的效率各有不同……48 3、rtnlb6突變株被農桿菌感染後,產生腫瘤及短暫表現T-DNA的效率增加……49 4、rtnlb7突變株被農桿菌感染後,產生腫瘤及短暫表現T-DNA的效率各有不同……49 五、阿拉伯芥中RTNLB1-21基因序列、信使RNA(messenger RNA/mRNA)序列、編碼區域序列(coding sequence/CDS)及蛋白質序列之同源關係分析………50 1、阿拉伯芥中RTNLB1-21基因序列同源關係分析………………50 2、阿拉伯芥中RTNLB1-21的信使RNA序列同源關係分析………51 3、阿拉伯芥中RTNLB1-21的編碼區域序列同源關係分析………51 4、阿拉伯芥中RTNLB1-21的蛋白質序列同源關係分析……52 六、利用Genevestigator分析RTNLB1-21基因在阿拉伯芥中不同組織部位中的表現情形…………………………………………………52 1、RTNLB1………………………………………………………………………52 2、RTNLB2………………………………………………………………………52 3、RTNLB3………………………………………………………………………53 4、RTNLB4……………………………………………………………………………53 5、RTNLB5……………………………………………………………………………53 6、RTNLB8……………………………………………………………………………53 7、RTNLB9…………………………………………………………………………….53 8、RTNLB10…………………………………………………………………………53 9、RTNLB11…………………………………………………………………………..53 10、RTNLB12…………………………………………………………………………54 11、RTNLB13…………………………………………………………………………54 12、RTNLB14…………………………………………………………………………54 13、RTNLB15…………………………………………………………………………54 14、RTNLB18…………………………………………………………………………54 15、RTNLB19…………………………………………………………………………54 16、RTNLB20…………………………………………………………………………55 17、RTNLB21…………………………………………………………………………55 七、RTNLB3、RTNLB5、RTNLB6及RTNLB7基因經由逆轉錄聚合酶連鎖反應(RT-PCR)結果顯示,在阿拉伯芥不同組織部位中表現各有不同…………………………55 1、RTNLB3……………………………………………………………………………56 2、RTNLB5……………………………………………………………………………56 3、RTNLB6……………………………………………………………………………56 4、RTNLB7……………………………………………………………………………56 八、ProRTNLB1::GUS轉殖株在阿拉伯芥各個生長階段及各器官中的表現情形…………..56 九、利用Genevestigator分析RTNLB在阿拉伯芥中受到不同生物性逆境、非生物性逆境、荷爾蒙或化學處理時的表現情形………………58 1、生物性逆境處理後RTNLB18至21基因的調控情形……………58 1.1、RTNLB18……………………………………………………………………..58 1.2、RTNLB19……………………………………………………………………..58 1.3、RTNLB20……………………………………………………………………..59 1.4、RTNLB21……………………………………………………………………..59 2、非生物性逆境處理後RTNLB18至21基因的調控情形……………59 2.1、RTNLB18………………………………………………………………………60 2.2、RTNLB19………………………………………………………………………60 2.3、RTNLB20………………………………………………………………………60 2.4、RTNLB21………………………………………………………………………60 3、賀爾蒙處理後RTNLB18至21基因的調控情形……………………60 3.1、RTNLB18………………………………………………………………………61 3.2、RTNLB19………………………………………………………………………61 3.3、RTNLB20………………………………………………………………………61 3.4、RTNLB21………………………………………………………………………61 4、化學逆境處理後RTNLB18至21基因的調控情形………………61 4.1、RTNLB18……………………………………………………………………..62 4.2、RTNLB19……………………………………………………………………..62 4.3、RTNLB20……………………………………………………………………..62 4.4、RTNLB21……………………………………………………………………...62 肆、討論………………………………………………………………64 一、RTNLB1~7基因於農桿菌感染時的功能分析………………………64 1、RTNLB1、2、4基因………………………………………64 2、RTNLB3、5、6及7基因……………………………………65 二、RTNLB1~21基因家族於植物生長階段可能扮演的角色分析……66 1、RTNLB1-7基因…………………………………………………67 2、RTNLB8-15基因…………………………………………………67 3、RTNLB16-21基因………………………………………………68 伍、參考文獻……………………………………………………………70 表一、阿拉伯芥中RTNLB基因序列、信使RNA(messenger RNA/mRNA)序列、編碼區域序列(coding sequence/CDS)及蛋白質序列之相關資料整理。……………………88 表二、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現的轉殖株之第二世代植株的分離率。…………………………… …………………………….91 表三、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現轉殖株的RTNLB2基因的RNA之累積量。…………………………………92 表四、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現轉殖株被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。……93 表五、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現的轉殖株之第二世代植株的分離率。………………………………………………94 表六、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現轉殖株的RTNLB1基因的RNA之累積量。…………………………………………95 表七、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現轉殖株被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。………………96 表八、T7-tagged RTNLB4大量表現的轉殖株之第二世代植株的分離率。……………97 表九、T7-tagged-RTNLB4大量表現轉殖株的RTNLB4基因的RNA之累積量。………98 表十、T7-tagged-RTNLB4大量表現轉殖株被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。…………………………………………99 表十一、阿拉伯芥中RTNLB基因之At編號及T-DNA插入突變株(T-DNA insertion mutants)。………………………………………………100 表十二、rtnlb3、rtnlb5、rtnlb6及rtnlb7的T-DNA insertion突變株之RTNLB3、RTNLB5、RTNLB6或RTNLB7基因的RNA之累積量。………102 表十三、rtnlb3、rtnlb5、rtnlb6及rtnlb7的T-DNA insertion突變株被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。………103 表十四、阿拉伯芥中RTNLB1~15基因於不同組織部位中的表現量。……104 表十五、阿拉伯芥中RTNLB18~21基因於不同組織部位中的表現量。……107 表十六、ProRTNLB1::GUS轉殖株之第二世代植株的分離率。………110 表十七、利用Genevestigator分析阿拉伯芥植株遭受生物性逆境、非生物性逆境、賀爾蒙或化學試劑處理後,RTNLB18~21基因之表現情形量化結果。…………………111 圖1、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現的轉殖植物的第二世代轉殖株的分離率結果。………………………………114 圖2、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現的轉殖植物內RTNLB2 RNA累積量之分析。……………………………………115 圖3、T7-tagged-RTNLB2大量表現的轉殖植物內T7-tagged-RTNLB2重組蛋白質累積量之分析。…………………………………………116 圖4、RTNLB2、T7-tagged-RTNLB2或His-tagged-RTNLB2大量表現的轉殖植物被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。……117 圖5、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現的轉殖植物的第二世代轉殖株的分離率結果。………………………………………118 圖6、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現的轉殖植物內RTNLB1 RNA累積量之分析。……………………………………………119 圖7、T7-tagged-RTNLB1大量表現的轉殖植物內T7-tagged-RTNLB1重組蛋白質累積量之分析。…………………………………………120 圖8、His-tagged-RTNLB1大量表現的轉殖植物內His-tagged-RTNLB1重組蛋白質累積量之分析。………………………………………121 圖9、T7-tagged-RTNLB1或His-tagged-RTNLB1大量表現的轉殖植物被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。………………122 圖10、T7-tagged-RTNLB4大量表現的轉殖植物的第二世代轉殖株的分離率結果。….123 圖11、T7-tagged-RTNLB4大量表現的轉殖植物內RTNLB4 RNA累積量之分析。......124 圖12、T7-tagged-RTNLB4大量表現的轉殖植物內T7-tagged-RTNLB4重組蛋白質累積量之分析。………………………………………………125 圖13、T7-tagged-RTNLB4大量表現的轉殖植物被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。……………………………………126 圖14、rtnlb3突變株內T-DNA插入植物染色體位置之分析。…………127 圖15、rtnlb3、rtnlb5、rtnlb6及rtnlb7的T-DNA insertion突變株之RTNLB3、RTNLB5、RTNLB6或RTNLB7基因的RNA之累積量分析。……128 圖16、rtnlb3、rtnlb5、rtnlb6及rtnlb7的T-DNA insertion突變株被土壤農桿菌感染後產生腫瘤及短暫表現T-DNA的效率。…………129 圖17、rtnlb5突變株內T-DNA插入植物染色體位置之分析。………130 圖18、rtnlb6突變株內T-DNA插入植物染色體位置之分析。………131 圖19、rtnlb7突變株內T-DNA插入植物染色體位置之分析。………132 圖20、阿拉伯芥的 RTNLB1-21 基因的同源關係樹狀圖(phylogenetic tree)。……133 圖21、阿拉伯芥的RTNLB1-21之信使RNA(mRNA)同源關係樹狀圖。……134 圖22、阿拉伯芥的RTNLB1-21編碼區域(coding sequence/CDS)的同源關係樹狀圖。…135 圖23、阿拉伯芥的RTNLB1-21之蛋白質同源關係樹狀圖。……………136 圖24、利用Genevestigator軟體分析 RTNLB1-15 在阿拉伯芥不同組織部位中的表現情形。……………………137 圖25、利用Genevestigator軟體分析 RTNLB18-21 在阿拉伯芥不同組織部位中的表現情形。…………………………………………138 圖26、逆轉錄聚合酶鍊式反應檢測RTNLB3、RTNLB5、RTNLB6及RTNLB7在花朵、花序、莖生葉、小苗、根及簇生葉的表現量。………………139 圖27、利用Genevestigator分析RTNLB1基因,在阿拉伯芥中不同組織部位中的表現情形。……………………140 圖28、ProRTNLB1::GUS的轉殖植物的第二世代轉殖株的分離率結果。……………141 圖29、ProRTNLB1::GUS-1 轉殖株各組織器官中的 GUS 蛋白質累積情形。………142 圖30、ProRTNLB1::GUS-2 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………143 圖31、ProRTNLB1::GUS-6 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………144 圖32、ProRTNLB1::GUS-22 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………145 圖33、ProRTNLB1::GUS-23 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………146 圖34、ProRTNLB1::GUS-36 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………147 圖35、ProRTNLB1::GUS-37 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………148 圖36、ProRTNLB1::GUS-40 轉殖株各組織器官中的 GUS 蛋白質累積情形。…………149 圖37、利用Genevestigator分析阿拉伯芥以生物性逆境處理後,RTNLB18~21基因之表現情形。……………………………………150 圖38、利用Genevestigator分析阿拉伯芥以非生物逆境處理後,RTNLB18~21基因之表現情形。………………………………………151 圖39、利用Genevestigator分析阿拉伯芥以賀爾蒙處理後,RTNLB18~21基因之表現情形。………………………………………152 圖40、利用Genevestigator分析阿拉伯芥以化學試劑處理後,RTNLB18~21基因之表現情形。…………………………………………153 附表一、引子序列表。……………………………………………………154 附圖1、用於產生大量表現RTNLB2轉殖株的質體製造圖解。…………156 附圖2、用於產生大量表現T7-tagged RTNLB1轉殖株的質體製造圖解。…………157 附圖3、用於產生大量表現T7-tagged RTNLB2轉殖株的質體製造圖解。…………158 附圖4、用於產生大量表現T7-tagged RTNLB4轉殖株的質體製造圖解。…………159 附圖5、用於產生大量表現His-tagged RTNLB1轉殖株的質體製造圖解。…………160 附圖6、用於產生大量表現His-tagged RTNLB2轉殖株的質體製造圖解。…………161zh_TW
dc.language.isoen_USzh_TW
dc.publisher生命科學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2610201014152400en_US
dc.subjectarabidopsisen_US
dc.subject阿拉伯芥zh_TW
dc.subjectagrobacteriumen_US
dc.subjectRTNLBen_US
dc.subject農桿菌zh_TW
dc.subjectRTNLBzh_TW
dc.title阿拉伯芥AtRTNLB1至AtRTNLB7基因家族於土壤農桿菌感染植物過程之功能分析zh_TW
dc.titleFunctional studies of the AtRTNLB1~ AtRTNLB7 genes in the Agrobacterium-mediated plant transformation processen_US
dc.typeThesis and Dissertationzh_TW
item.languageiso639-1en_US-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.grantfulltextnone-
item.fulltextno fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
Appears in Collections:生命科學系所
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