甜蛋白Monellin在巴斯德毕赤氏酵母中的胞内表达  

Expression of Monellin, a Sweet Protein, in Pichia pastoris

作　　者：于学红[1] 袁汉英[1] 高卜渝[1] 范长胜[1] 周欣[1] 李育阳[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所,遗传工程国家重点实验室,上海200433

出　　处：《复旦学报（自然科学版）》2002年第6期650-654,共5页Journal of Fudan University：Natural Science

摘　　要：将PCR扩增得到的Monellin基因片段插入Pichiapastoris胞内表达质粒pPIC3.5K',然后转入Pichiapas torisSMD1168受体菌中.用G418筛选高拷贝菌株,并进一步对其进行PCR鉴定.获得的阳性克隆菌株经甲醇诱导60h,SDS PAGE结果显示菌株破壁液中含有表达的Monellin,其大小约为11ku,并具有甜度.The gene, monellin, encoding a single chain Monellin was cloned into Pichia pastoris expression vector pPIC3.5k'. Linearized pPIC3.5k'/monellin was transformed into P. pastoris SMD 1168. Recombinants with the highest resistance to G418 were selected and then identified by PCR analysis. The selected tranformants were cultivated in flasks. The expressed Monellin with 11 ku in size could be detected on SDSPAGE after 60 hours induction with methanol. The extracts of cultured yeast and the purified products have a strong taste of sweetness.

关 键 词：甜蛋白 MONELLIN 巴斯德毕赤氏酵母 酵母表达 

分 类 号：Q51[生物学—生物化学] Q786