摘要
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（Ｌａｔｅｏｌａｂｒａｘｊａｐｏｎｉｃｕｓ）生长激素基因克隆到酵母整合型质粒载体ｐＨＩＬＤ２，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，ＳＤＳＰＡＧＥ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 pHILD2, which was then transformed into his4 mutant yeast GS115. PCR fast detection methods was performed to screen the positive transformants and dotblotting was used to screen the multiplecopy transformants. Via inducing AOX1 promoter by methanol, growth hormone could be expressed intracellularly. SDSPAGE and Western blot were applied to confirm the product.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1998年第2期140-143,共4页
Progress In Biochemistry and Biophysics
关键词
鲈鱼
生长激素
胞内表达
甲醇酵母
Lateolabrax japonicus growth hormone, Pichia pastoris, intracellular expression
