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 SDSPAGE after 60 hours induction with methanol. The extracts of cultured yeast and the purified products have a strong taste of sweetness.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
2002年第6期650-654,共5页
Journal of Fudan University：Natural Science
