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題 名 | Cloning and Characterization of Insulin-Like Growth Factor I cDNA from Black Seabream (Acanthopagrus Schlegeli)=黑鯛類胰島素生長因子基因之選殖與定序 |
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作 者 | 陳泓志; 林錦華; 龔紘毅; 李秋月; 張繼堯; 陳鐵雄; 吳金洌; | 書刊名 | 動物研究學刊 |
卷 期 | 37:3 1998.07[民87.07] |
頁 次 | 頁213-221 |
分類號 | 387.1015 |
關鍵詞 | 類胰島素生長因子; 鯛科; 互補去氧核糖核酸; IGF-1; Sparidae; cDNA; |
語 文 | 英文(English) |
中文摘要 | 由於肝臟是 IGF 基因表現最多的部位,本研究擬由黑鯛肝臟組織,建立 Uni-ZAP XR cDNA 基因庫, 並進行 IGF 基因家族之選殖。 本實驗利用鱒魚 IGF-I cDNA 的 C 和 E 區,設計引子,以反轉錄聚合�t連鎖反應來合成黑鯛 IGF-I cDNA 探針,以此內生性探針自 黑鯛肝臟 cDNA 基因庫中篩選出 IGF-I cDNA 基因。 以限制�t圖譜分析, 篩選到的十六個選殖株, 整理為大小自 1.8 至 2.3 kb 的六種 cDNA 片斷之選殖株, 其中兩個較長的 IGF-I cDNA 基因, 經定序分析後,大小分別為 2238 bp 及 2299 pb,發現這兩個基因除了一個在 5' 端少了 61 個核��酸序列和在 3' 端非轉譯區 發現六個核��酸不同,其餘序列皆相同,推測是一種多型態( polymorphism )現象或基因 雙倍化( duplication )所形成的。 黑鯛 IGF-I cDNA 可轉譯出 185 個胺基酸,其中包含 44 個胺基酸的訊息胜�鞳A67 個胺基 酸含 B、C、A、D 區的成熟胜�鞳A以及 74 個胺基酸的 E 區,而原胜�� IGF-I 由 B、C、A 、D、和 E 區所組成。 其中值得注意的是已知物種的 IGF-I 成熟胜�颿O留性相當高,比較 黑鯛與其它魚類 IGF-I 胺基酸序列分別為鯛,鮭, 鱒,鯉,鯰,盲鰻和文昌魚有 100%, 91% ,91%,87%,87,58%,和 48% 的同源性。而黑鯛 IGF-I 的 E 區相較於鮭科魚類是屬 於最長的,故此選殖出之黑鯛 IGF-I cDNA 基因應屬 IGF-I Ea-4 家族而且序列比對符合現 行之魚類種源關係圖,是為鯛科。 |
英文摘要 | A cDNA library was constructed in Uni-ZAP XR using mRNA from the liver of black seabream, Acanthopagrus schlegeli. In this study, we designed a pair of primers from the C and E domains of trout IGF-I cDNA, and synthesized an internal probe of IGF-I from the liver of black seabream by reverse transcription/polymerase chain reaction (RT/PCR). Using the internal probe to screen the cDNA library, we obtained 16 positive clones. Subsequent restriction enzyme map analysis suggested that about 6 forms ranging in size from 1.8 kilo-base pair (kb) to 2.3 kb of IGF-I were present. We took 2 longer cDNA clones to read the full sequence. Sequences of 2 clones of IGF-I cDNAs were found to be 2238 base pairs (bp) and 2299 bp in length. Except for a sequence of 61 nucleotides missing in the 5'-untranslated region (5'-UTR) and 6 nucleotides being different in the 3'-untranslated region (3'-UTR), the other nucleotides of these 2 clones are identical. This showed that the black seabream IGF-I gene contains polymorphism or gene duplication. Both cDNAs containing an open reading frame (ORF) encode 185 amino acids, including a 44-amino acids leader peptide, the 67-amino acids mature peptide in the B, C, A, and D domains, and a 74-amino acids extended carboxyl-terminal peptide in the E domain. The predicted propeptide of IGF-I can be divided into B, C, A, D, and E domains. Owing to the conserved mature peptide of IGF-I, we compared IGF-I of other teleosts with that of black seabream. The comparisons showed 100%, 91%, 91%, 87%, 87%, 58%, and 48% amino acid identity, respectively, with the IGF-I of Sparus, salmon, trout, carp, catfish, hagfish, and amphioxus. Moreover, we compared the E domain of IGF-I of Salmonidae with that of black seabream. The comparisons revealed that the E domain of black seabream belongs to the largest Ea from. These data imply that black seabream cDNAs encode a particular subtype of IGF-I from liver, IGF-I Ea-4. Furthermore, this IGF-I is phylogenetically most closely related to that of Sparidae. |
本系統中英文摘要資訊取自各篇刊載內容。