Position: Home > Articles > Expression of Monellin-EGFP Fusion Protein in Pichia pastoris
FOOD SCIENCE
2012
(13)
171-175
Monellin-EGFP融合蛋白在毕赤酵母中的表达
作 者：
金筱耘;赵爱春;李军;李镇刚;王茜龄;吴存容;余茂德
单 位：
西南大学生物技术学院
关键词：
Monellin甜蛋白;绿色荧光蛋白;真核表达;桑树
摘 要：
根据已报道的Monellin甜蛋白氨基酸序列,结合毕赤酵母与桑树密码子的偏好性,人工设计合成单链monellin基因,并与增强型绿色荧光蛋白EGFP基因连接,构建融合表达载体。采用电击转化法成功将表达载体导入毕赤酵母GS115中,并获得高效表达的重组子pPIC9K-M-E。经PCR检测,SDS-PAGE和Western Blot杂交证实已表达出融合蛋白,该蛋白经纯化与盲测实验,结果显示其甜度约为标准蔗糖的500倍。
译 名：
Expression of Monellin-EGFP Fusion Protein in Pichia pastoris
作 者：
JIN Xiao-yun,ZHAO Ai-chun,LI Jun,LI Zhen-gang,WANG Xi-ling,WU Cun-rong,YU Mao-de*(College of Biotechnology,Southwest University,Chongqing 400716,China)
关键词：
monellin sweet protein;EGFP;eukaryotic expression;mulberry
摘 要：
According to the reported amino acid sequence of single-chain monellin,the codon bias of Pichia pastoris and Morus alba L.,a synthetic monellin gene was achieved.Then,the enhanced green fluorescent protein(EGFP) gene was added into the downstream of monellin gene to construct an expression vector named as pPIC9K-M-E.Based on transformation of yeast by electroporation,the expression vector pPIC9K-M-E was successfully imported into Pichia pastoris.PCR amplification,fluorescence detection,Western Blot analysis and SDS-PAGE results demonstrated that the fusion protein was expressed and the fusion protein revealed high intensity of sweetness,which was almost 500 times sweeter than the sucrose with same amount.
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