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頁籤選單縮合
題名 | Generation of Transgenic Tilapia by using Cryopreserved Electroporated Sperm to Mediate Gene Transfer=利用冷凍保存精子做基因載體製作基因轉殖吳郭魚 |
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作者 | 陳昭德; 蔡添財; 施志民; 陳榮華; Chen, Jau-der; Tsay, Tian-tsair; Shie, Jei-min; Chern, Rong-hwa; |
期刊 | 臺灣水產學會刊 |
出版日期 | 19981200 |
卷期 | 25:4 1998.12[民87.12] |
頁次 | 頁265-272 |
分類號 | 439.5 |
語文 | eng |
關鍵詞 | 精液冷凍保存; 電穿孔法; 基因轉殖吳郭魚; Cryopreservation; Electroporation; Transgenic tilapia; |
中文摘要 | 吳郭魚精液先以精虫抑制活化緩衝液清洗處理後,再懸浮於 0.85% 淡水魚生理食 鹽水中,用於電穿孔處理時做為轉殖基因的接受者。這些電穿孔處理過帶有轉殖基因的精子 ,立刻以液態氮冷凍保存達 40 天。解凍後的精子與成熟吳郭魚卵授精,解凍後的精子其孵 化率為 23.79% 較用新鮮的精子授精的孵化率 43.25% 為差。解凍後的精子細胞,內含轉殖 基因可利用南方氏核��酸雜交法檢測到。又授精後孵化的小魚,於萃取核��酸做聚合�t連鎖 反應法檢測,亦可証實轉殖基因的存在,因此冷凍保存電穿孔處理過帶有轉殖基因的精子, 可做為轉殖基因的攜帶者直接用於人工授精製作基因轉殖吳郭魚。 |
英文摘要 | Tilapia milt treated with sperm deactivator and supended in 0.85% freshwater fish saline was used as the DNA recipient for gene transfer by electroporation. These electroporated sperm were immediately cryopreserved in liquid nitrogen and stored for 40 days. As measured by hatching percentages, the viability of thawed cryopreserved sperm used for fertilization was 23.79% less than that 43.25% of fresh sperm. The presence of the transgene within the post-thawed electroporated sperm was confirmed by Southern blot, and the presence of the transgene in the genome of transgenic fry was confirmed by PCR. We conclude that cryopreserved electoporated tilapia sperm is a good transgene mediator that can be used to generate transgenic tilapia. |
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