鲈鱼生长激素在甲醇酵母中的胞内表达

Intracellular Expression of Lateolabrox japonicus Growth Hormone in Methyltrophic Yeast, Pichia pastoris.

甲醇酵母ｐｉｃｈｉａｐａｓｔｏｒｉｓ是一种理想的真核蛋白高水平表达系统．将鲈鱼（Ｌａｔｅｏｌａｂｒａｘｊａｐｏｎｉｃｕｓ）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．

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.