重组人Endostatin在甲醇酵母Pichia Pastoris中的分泌表达

The Secretion and Expression of Recombinant Human Endostatin in Methanol Yeast Pichia Pastoris

为了获得高效表达分泌型重组人Endostatin菌株 ,利用基因工程技术 ,将目的基因连接到Pichia表达载体 pPIC9K中 ,将该重组质粒转入GS115菌株后筛选His阳性菌株 ,并用G4 18筛选高拷贝阳性克隆菌 .重组表达的Endostatin经SDS PAGE电泳 ,可在分子量 2 2kD处见到单一目标带 .经体外生物学活性检测 ,Pichia菌株表达的Endostatin能有效地抑制培养的ECV30 4细胞及P6 1细胞的生长 .

In order to get secreting methanol yeast strain of human Endoatatin(EN) with high expression, using gene recombination technique, the authors combinate the En gene into vector pPIC9k and transfecte EN-pPIC9k to yeast strain GS115. After screening, some positive strains were got. Which can secrete EN. The supernatant fluid were performed by SDS-PAGE, a single band was seen at the position of 22kD or so. The expressed EN can effectively inhibits the growth of cultured cells ECV304 and P61.