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來源資料
頁籤選單縮合
題名 | Vertical Transmission of Transgene from Transgenic Founder to Their Offspring Showing a Phenomenon of Losing Transgene from the Genomes of Transgenic Tilapia, Oreochromis niloticus=基因轉殖吳郭魚轉殖基因垂直遺傳至子代顯現嵌入染色體中轉殖基因脫落的現象 |
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作者 | 陳昭德; 蔡添財; 陳榮華; 施治民; 湯雅婷; 李芷懿; 李佳育; | 書刊名 | 臺灣水產學會刊 |
卷期 | 27:4 2000.12[民89.12] |
頁次 | 頁283-294 |
分類號 | 439.5 |
關鍵詞 | 轉殖基因排斥; 聚核酶連鎖反應法; 基因轉殖吳郭魚; Transgenic gene exclusion; PCR; Transgenic tilapia; |
語文 | 英文(English) |
中文摘要 | 以精子做載體攜帶轉殖基因,再與成熟卵受精孵化成基因轉殖吳郭,利用轉殖基因 中JAK起動子及黃鰭鯛促成長激素之唯一連結核茹酸片段不存在於吳郭魚(Oreochromis niloticus)染色體上,以聚核?連鎖反應法快速檢定轉殖基因是否嵌入基因轉殖吳郭魚染色體 中。結果顯示轉殖基因不僅傳入子一代且已傳入子二代。聚核?連鎖反應法檢定之基因轉殖 吳郭魚,抽取去氧核醣核茹酸(DNA)作南方氏轉漬雜交(Southern Hybridization),証明轉殖基 因存在於染色體中。由肝臟抽取核核茹酸(RNA)作北方氏轉漬雜交(Northern Hybridization), 証明轉殖基因在轉錄(Transcription)層次有基因表現。由基因轉殖吳郭魚的血清作西方氏免疫 雜交(Western Blot)檢測,部份基因轉殖吳郭魚可檢測到轉殖基因所表現的促成長激素。但在 追蹤轉殖基因是否穩定嵌入吳郭魚染色體中,在第一年內皆可追蹤到轉殖基因的存在,於蓄 養兩年後,不論是子一代或子二代,轉殖基因皆因宿主染色體的排斥或不明原因脫落而消失。 |
英文摘要 | Sperm that electroporated by transgene consisting of JAK promoter plus ypGH were fertilized by mixing with mature eggs to generate transgenic tilapias (Oreochromis niloticus). The presence of transgene within the gnome of transgenic tilapia was detected by PCR analysis of the fusion components of JAK promoter plus ypGH. The expression of ypGH product was detected by RT-PCR at transcriptional level and immunoblotted by a specific anti-ypGH antibody at translational level. The presence of transgene was identified by PCR analysis of the most DNA samples at younger fishes that generated within the first year since development. However, the situation of deletion of transgenes from the genomes of the identified transgenic tilapias was detected during the following farming period and the transgene was almost completely lost after two years rearing. This phenomenon of unstable integration of transgene was observed by examining the genomes of transgenic founder, generation F1 and F2 during the past four years' farming. |
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