DC 欄位 | 值 | 語言 |
dc.contributor | 繆希椿 | zh-TW |
dc.contributor | Miaw, Shi-Chuen | en |
dc.contributor | 臺灣大學:免疫學研究所 | zh-TW |
dc.contributor.author | 楊馥瑋 | zh-TW |
dc.contributor.author | Yang, Fu-Wei | en |
dc.creator | 楊馥瑋 | zh-TW |
dc.creator | Yang, Fu-Wei | en |
dc.date | 2009 | en |
dc.date.accessioned | 2010-05-10T10:52:04Z | - |
dc.date.accessioned | 2018-07-09T01:39:18Z | - |
dc.date.available | 2010-05-10T10:52:04Z | - |
dc.date.available | 2018-07-09T01:39:18Z | - |
dc.date.issued | 2009 | - |
dc.identifier.other | U0001-1708200917402600 | en |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/181809 | - |
dc.description.abstract | c-Maf是一個b-Zip家族的轉錄因子,根據之前的文獻報導,c-Maf在Th2細胞中對IL-4基因調控是一個必要的存在。而最近的文獻則是報導 c-Maf與 IL-21在Th17細胞中的關係。這篇文獻發現在c-Maf KO小鼠中,IL-21的基因表現是有缺陷的,也就是說,c-Maf很有可能是調控IL-21基因表現的一個上游分子。由於c-Maf本身是一個轉錄分子,因此我們假設c-Maf會調控IL-21基因表現。為了確定c-Maf調控IL-21基因是經由直接接觸IL-21 gene locus或是透過與其它不明轉錄蛋白作用再影響到IL-21基因表現,我們利用電腦軟體在IL-21 promoter的基因序列中找尋MARE。我們在-1.3kb中總共找到兩個half MARE和兩個V-MARE。為了確定c-Maf會藉由哪一個MARE影響IL-21的表現,我們建構了具有點突變或是截短的promoter片段的pGL3質體,再利用這些質體進行Luciferase assay。我們的研究結果指出c-Maf最有與位在-192 ~ -205 kb的V-MARE作用,進而影響IL-21基因表現。除了以上的實驗,我們也去測試SUMOylation會不會讓c-Maf對IL-21基因表現造成影響。在這方面,我們的結論是:c-Maf的轉錄能力在調控IL-21基因系統中,不受到SUMOylation的影響。 | zh-TW |
dc.description.abstract | c-Maf is a b-Zip transcription factor, which was found to be an essential role in regulating IL-4 expression in Th2 cells. Recent study has investigated the relationship between c-Maf and IL-21 gene expression in Th17 cells. Their study uncovered shortage of IL-21 production in c-Maf KO mice, which implied that c-Maf might be an upstream molecule of IL-21. Since c-Maf itself is a transcription factor, we hypothesized that c-Maf should regulate IL-21 gene expression. To identify whether c-Maf regulates IL-21 expression by directly or indirectly binding on IL-21 promoter, we went to investigate MARE sequence on IL-21 promoter. We found two half MARE and two V-MARE in -1.3kb upstream region. We constructed several point mutatated and truncated promoter fragments onto pGL3 Basic vector, and performed luciferase assay. Our results pointed out that the putative c-Maf binding site should be V-MARE locating -192 ~ -205kb. On the other hand, the SUMOylation effect of c-Maf on IL-21 gene expression was tested. We concluded that c-Maf transactivation ability on IL-21 gene expression was not affected by SUMOylation. | en |
dc.description.tableofcontents | Ackowledgement…………………………………...………………….ibstract..………………………………………………….....................ii 要………………………………………………………………….. iiihapter I Bckground.………….…………………….................. 1.1 The characteristic cytokine expression of T helper cells….……... 1.2 Th2 cells differentiation……………………..…….…………. 2.3 Th17 cells differentiation…………………...………………... 3.4 An overview of c-Maf and its function…………………. 4.5 Post-translation modification of c-Maf…………………….…… 6.6 IL-21……………………………………………………….… 7. Rationale…………………………………………………….…… 8. Hypothesis……………………………………….... 9hapter II Methods……………………...………… 10.1 Cell Culture…………………………………………………… 10.2 Plasmids……………………………………………………………. 10.3 Transfection………………………………………………………....13.4 Luciferase assay………………………………….………………….13.5 Western Blot………………………………………………………...13.6 Statistics……………………………………………………………..14hapter III Experimental Materials................................ 15.1 Cell line……………………………………………………………..15.2 Antibodies.…………………………………………………………..15.3 Enzymes…………………………………………………………….15.4 Kits………………………………………………………………….16.5 Chemicals and reagents……………………………………………..16hapter IV Results..…………………….….…………………….19.1 c-Maf induce murine IL-21 gene expression………………………..19.2 Putative c-Maf binding sites may locate on -250bp romoter region…………………………………….……………….20.3 The binding site of wt c-Maf is at V-MARE site locating 192 ~ -205………………………………………………………….21.4 Sumoylation of c-Maf has no significant impact on IL-21 expression..………………………….……………………...22hapter V Discussion…...……………………………..…………24eference………………………………………..………………….....28hapter VI Experimental Figures…...…..…………… 32ig.1 The map of pGL3-Basic mIL-21 promoter constructs… 33ig.2 c-Maf is able to transactivate mIL-21 expression…................. 34ig.3 Two promoter deletion constructs designation……………………35ig.4 Slightly decreased transactivity of wt c-Maf on two eletion mutants…………………………………………………..36ig.5 Three MARE mutants'' designation………………………. 37ig.6 Slightly decreased transactivity of wt c-Maf on two ARE mutants..…...…………………………………….……….38ig.7 Parallel tendency of two wt c-Maf dosages on our mutants………………………………………….…………….39ig.8 wt c-Maf failed to induce pGL3 mIL-21 promtoer 200 expression…………..…………………………...…………..40ig.9 DeSumoylated environment lead to upregulated transactivity of t c-Maf on mIL-21 expression……………………..………..….43ig.10 wt c-Maf isn’t related to Sumoylation in mIL-21 xpression system……………………………………………….44hapter VII Supplementary Data…………………………………..45Fig.1 Slightly decreased transactivity of wt c-Maf on two eletion mutants…………………………………………………..46Fig.2 Slightly decreased transactivity of wt c-Maf on two ARE mutants…………………………………………… 48Fig.3 Parallel tendency of two wt c-Maf dosages on our mutants………………………………………………………50Fig.4 wt c-Maf failed to induce pGL3 mIL-21 promtoer 200 expression………………………………………………….53 | en |
dc.format | application/pdf | en |
dc.format.extent | 2317361 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | en | en |
dc.language.iso | en_US | - |
dc.subject | 小鼠 | zh-TW |
dc.subject | c-Maf | en |
dc.subject | IL-21 | en |
dc.subject | Th17 | en |
dc.title | c-Maf在調控小鼠IL-21基因所扮演的角色 | zh-TW |
dc.title | Regulation of murine IL-21 gene by c-Maf | en |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/181809/1/ntu-98-R96449002-1.pdf | - |
item.languageiso639-1 | en_US | - |
item.grantfulltext | open | - |
item.fulltext | with fulltext | - |
顯示於: | 免疫學研究所
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