| 鲈鱼生长激素在甲醇酵母中的胞内表达 |
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| 引用本文： | 陈丹,杨丰,王玮,徐洵.鲈鱼生长激素在甲醇酵母中的胞内表达[J].生物化学与生物物理进展,1998,25(2):140-143. |
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| 作者姓名： | 陈丹 杨丰 王玮 徐洵 |
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| 作者单位： | 国家海洋局第三海洋研究所，厦门 361005;国家海洋局第三海洋研究所，厦门 361005;国家海洋局第三海洋研究所，厦门 361005;国家海洋局第三海洋研究所，厦门 361005 |
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| 摘 要： | 甲醇酵母pichia pastoris是一种理想的真核蛋白高水平表达系统.将鲈鱼(Lateolabrax japonicus)生长激素基因克隆到酵母整合型质粒载体pHIL-D2，经转化his4缺陷型酵母GS115，用PCR方法筛选阳性转化子，并用斑点印迹法筛选多拷贝转化子，经甲醇诱导表达，SDS-PAGE和蛋白质印迹杂交结果证实了表达产物为重组的鲈鱼生长激素.
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| 关 键 词： | 鲈鱼生长激素，pichia pastoris，胞内表达 |
| 收稿时间： | 1997/1/29 0:00:00 |
| 修稿时间： | 8/4/1997 12:00:00 AM |
Intracellular Expression of Lateolabrox japonicus Growth Hormone in Methyltrophic Yeast, Pichia pastoris |
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| CHEN Dan,YANG Feng,WANG Wei and XU Xun.Intracellular Expression of Lateolabrox japonicus Growth Hormone in Methyltrophic Yeast, Pichia pastoris[J].Progress In Biochemistry and Biophysics,1998,25(2):140-143. |
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| Authors: | CHEN Dan YANG Feng WANG Wei and XU Xun |
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| Institution: | The 3rd Institute of Oceanography, State Oceanic Administration (SOA), Xiamen 361005, China;The 3rd Institute of Oceanography, State Oceanic Administration (SOA), Xiamen 361005, China;The 3rd Institute of Oceanography, State Oceanic Administration (SOA), Xiamen 361005, China;The 3rd Institute of Oceanography, State Oceanic Administration (SOA), Xiamen 361005, China |
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| Abstract: | The methylotrophic yeast, Pichia pastoris heterologous gene expression system was utilized to produce attractive levels of a variety of intracellular and extracellular proteins of interest. Lateolabrox japonicus growth hormone gene was cloned into the yeast integrative vector pHIL-D2, which was then transformed into his4 mutant yeast GS115. PCR fast detection methods was performed to screen the positive transformants and dot-blotting was used to screen the multiple-copy transformants. Via inducing AOX1 promoter by methanol, growth hormone could be expressed intracellularly. SDS-PAGE and Western blot were applied to confirm the product. |
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| Keywords: | Lateolabrax japonicus growth hormone Pichia pastoris intracellular expression |
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