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題名 | Genomic Structure and Promoter Region of C-fos Gene of Round-Spotted Pufferfish, Tetraodon Nigroviridis (Syn. T. Fluviatilis)=圓斑河豚(Tetraodon nigroviridis [Syn. T. fluviatilis]) c-fos基因結構與起動力活性分析 |
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作 者 | 張茂山; 呂健宏; 姚振文; 黃火鍊; 黃銓珍; | 書刊名 | 動物研究學刊 |
卷期 | 36:3 1997.07[民86.07] |
頁次 | 頁159-169 |
分類號 | 388.5 |
關鍵詞 | 基因結構; 起動子; 圓斑河豚; Genomic DNA sequence; C-fos; Round-spotted pufferfish tetraodon nigroviridis; Chloramphenicol acetyltransferase; CAT; |
語文 | 英文(English) |
中文摘要 | 圓斑河豚屬於豚形目四齒豚科,廣泛分佈於東南亞及東印度的河口區域,在台灣 容易自水族館購得,其基因體大小約為380Mb,只有人類基因體大小的八分之一,適於從 事基因體結構分析與比較。 在鯉魚蛋白激 JAK1基因的啟動子區域有兩處轉錄因子AP1結合的核酸序列,為了研 究AP1如何調控JAK1基因的轉錄,我們便著手魚類c-fos基因的分子選殖。在本報告中, 我們自圓斑河豚染色基因體積因庫篩選得到c-fos基因並定其基因結構。實驗的結果顯示圓 斑河豚c-fos基因共包含4個外顯子和3個內隱子,可轉譯一個含374個胺基酸的蛋白質, 其分子量約為42kb,其胺基酸序列與虎河豚(Fugu rubripes)、人類、小白鼠與雞的相同度為 90%、58%、58%與53%。我們利用引子延展分析決定了兩個可能的轉錄起始點,並且在圓 斑河豚c-fos基因0.75kb的啟動子區域中也發現了CREB、E2A、Ets-1、GATA-1、HNF-5、 c-Myb及Sp1等轉錄因子的核酸結合序列。另外在氯徽素乙醯轉換酵素活性分析的實驗中, 我們發現在圓斑河豚c-fos基因-746到+153的啟動子區域具有較對照組pRSV-CAT高出2.7 倍的氯徽素乙醯轉換酵素活性,而此啟動子區域在5%胎牛血清的刺激下,其酵素活性亦明 顯增加,證明圓斑河豚c-fos基因-746到+153的區域是一有功能的啟動子區域。 |
英文摘要 | Mau-Sun Chang, Jiann-Horng Leu, Chen-Wen Yao, Fore-Lin Huang and Chang-Jen Huang (1997) Genomic structure and promoter region of c-fos gene of round-spotted pufferfish, Tetraodon nigroviridis (Syn. T. fluviatilis). Zoological Studies 36(3): 159-169. The round-spotted pufferfish c-fos gene was isolated form a liver genomic library. Complete DNA sequences, including 0.75-kb of the promoter region were dertermined. Our data indicate that this gene is composed of 4 exons and 3 introns spanning about 3kb. This gene encodes a protein of 374 amino acids with a molecular mass of 42 262 Da. The amino acid sequence identities between this puffer fish c-fos protein and fugu (another pufferfish species), human, mouse, and chicken c-fos protein are 90%, 58%, 58% and 53%, respectively. Two potential transcription initiation sites were located by primer extension analysis. Examination of 0.75-kb of a 5'-flanking sequence revealed potential binding sites for a variety of transcription factors such as CREB, E2A, Ets-1,GaTA-1, HNF-5, c-Myb, and Sp1. When the 0.9-kb DNA fragment (-746 to +153) was placed upstream of the chloramphenicol acetyl- transferase (CAT) reporter gene and transfected into a carp CF cell line, it could drive the synthesis of CAT enzyme 2.7 times more efficiently than could the RSV promoter. Although lacking the known serum-response element, the CAT activity or this region still could be induced by serum. |
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