鲈鱼生长激素在甲醇酵母中的胞内表达
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Intracellular Expression of Lateolabrox japonicus Growth Hormone in Methyltrophic Yeast, Pichia pastoris
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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.

    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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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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  • 收稿日期:1997-01-29
  • 最后修改日期:1997-08-04
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