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題名 | 心臟型脂肪酸結合蛋白基因剔除小鼠胚胎幹細胞株之選殖=Cloning the Mouse Embryo Stem Cells with Knock-out Heart-type Fatty Acid Binding Protein (H-FABP) Gene |
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作者 | 楊媖琁; 張秀鑾; 劉世華; 吳美莉; 李坤雄; 莊秀琪; Yang, Ying-hsuan; Chang, Hsiu-luan; Liu, Shih-hwa; Wu, Mei-li; Li, Kwun-hsung; Chaung, Hso-chi; |
期刊 | 臺灣獸醫學雜誌 |
出版日期 | 20091200 |
卷期 | 35:4 2009.12[民98.12] |
頁次 | 頁331-342 |
分類號 | 437.2 |
語文 | chi |
關鍵詞 | 心臟型脂肪結合蛋白; 基因剔除; 胚胎幹細胞; 小鼠; Heart-type fatty acid binding protein; Gene targeting; Embryo stem cells; Mice; |
中文摘要 | 心臟型脂肪結合蛋白(H-FABP)存在於高脂肪需求之組織,尤其是心臟,作為吸收及運送的脂肪酸結合蛋白,其功能為促進運送長鏈脂肪酸至細胞內特定位置進行代謝,具有調節細胞內脂肪酸代謝之作用。本試驗應用基因重組技術以基因剔除(gene knockout)載體,於GT-N28 載體中連結包含兩段小鼠H-FABP基因之兩段同源性基因重組區域,以及neo(superscript R)及herpes simplex virus-1 thymidine kinase (HSV-TK)作為篩選基因產生H-FABP標的基因剔除載體,將該載體轉殖入小鼠129/svj品系之胚胎幹細胞培養,經確認其染色體DNA中含有剔除載體中H-FABP與neo(superscript R)之連結核酸片段,顯示篩選所得之胚幹細胞株已完成剔除載體與染色體標的基因之重組。 |
英文摘要 | The heart-type fatty acid binding protein (H-FABP) is primarily present in tissues with a high demand for fatty acids, especially in heart, to facilitate the uptake of long chain fatty acids and promote their intracellular trafficking to sites of metabolic conversions, and therefore modulate the intracellular fatty acid metabolism. Gene targeting in mouse embryonic stem (ES) cells is a fundamental methodology for generating mice with precise genetic modifications. In the present study, the H-FABP gene targeting vector was constructed by ligating two homologous regions of mouse HFABP gene, neo1 gene and HSV-TK gene into a pGT-N28 vector. The H-FABP gene targeting vector was then transfected into the mouse 129/svj embryo stem cells. The presence of a DNA fragment of partial H-FABP gene targeting vector ligating with neo1 gene confirmed the occurring recombination between H-FABP gene targeting vector and chromosomal homologous genes in transfected stem cells. |
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