臺灣博碩士論文加值系統

介白素20 (Interleukin-20；IL-20)屬於IL-10家族成員之一，此家族成員包含了IL-10、IL-19、IL-20、IL-22、IL-24及IL-26。在人體細胞內，IL-20位於染色體1q32上，包含了5個exon及4個intron。IL-20是一多功能性的細胞激素，包含有發炎、血管新生及趨化性 (chemoattractive)等功能。但IL-20的詳細生物功能依然不清楚。在本研究中我們想要去探討人類IL-20的基因表現。經由分析58位正常人和140位全身性紅斑性狼瘡 (system lupus erythematosus；SLE)病人的IL-20啟動子區域核酸序列，我們發現正常人或病人約10%的人，其IL-20啟動子上核酸序列-168/-169 (相對於轉錄起始點+1)間存在著一段318 bp的insertion-deletion polymorphism。經由啟動子活性 (promoter activity)分析，含有318 bp片段的重組基因啟動子活性比不含有318 bp片段的活性低，因此我們推測這318 bp是一個負向調控的序列。進一步，將這318 bp分成不同片段構築至含有luciferase報導基因 (reporter gene )的pGL3 basic vector，並轉染 (transfect)至HEK 293細胞中進行啟動子活性分析，得到核酸序列244-318是重要的區域。為了更進一步去分析核酸序列244-318上有哪些轉錄因子會和它結合便將核酸序列244-318分成不同的片段去進行electrophoresis mobility shifting assay (EMSA)分析，最後我們推測核酸序列267-271和277-286上有轉錄因子結合的序列。在本研究，我們推測負向調控轉錄因子經由與核酸序列267-271和277-286結合，導致IL-20啟動子活性降低，至於更詳細的機制仍需要更多的研究。

Interleukin-20 (IL-20) belongs to IL-10 family, including IL-10, IL-19, IL-20, IL-22, IL-24, and IL-26. In human, IL-20 locates on chromosome 1q32, and contains five exons and four introns. IL-20 is a pleiotropic cytokine with potent inflammatory, angiogenic, and chemoattractive characteristics. The detailed biological function of IL-20 is not very clear. In the study, we aimed to investigate the regulation of human IL-20 by analyzing its promoter region. After sequencing the IL-20 promoter from 198 individuals, including 58 normals and 140 systemic lupus erythematosus (SLE) patients, we discovered that there was a 318 bp-insertion-deletion polymorphism between -168 and -169 (related to the transcription initiation site +1). Insertion of the 318 bp in the promoter region decreased the promoter activity in all the fusion genes linked to pGL3 basic vector, suggesting that the region contains a negative regulatory element. Furthermore, different deletion constructs of the 318 bp in the fusion gene (pE) linked to pGL3 basic vector showed that the region from nucleotide sequence 244-318 was a critical region of the 318 bp. To identify the critical region with transcription factors, electrophoresis mobility shift assay (EMSA) was performed, revealing that the nucleotide sequence 272-291 has potential binding sites for the unidentified transcription factors. Further EMSA analysis showed the nucleotide sequence 267-271 and 277-286 might be the regions for binding of transcription factors. The binding of the unidentified transcription factors to these two regions might contribute to the suppressive activity of the 318 bp in IL-20 promoter.

誌謝 1
中文摘要 2
英文摘要 3
目錄 5
圖目錄 6
表目錄 7
附錄目錄 8
縮寫檢索表 9
儀器 10
第一章 緒論 12
第二章 實驗方法與材料 15
第三章 結果 30
第四章 討論 33
參考文獻 35
圖 40
表 53
附錄 55
自述 58

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