Identification and characterization of novel promoters for recombinant protein production in yeast Pichia pastoris

被引:32
|
作者
Xu, Ning [1 ]
Zhu, Jinxiang [1 ]
Zhu, Qiaoyun [1 ]
Xing, Yanzi [1 ]
Cai, Menghao [1 ]
Jiang, Tianyi [3 ]
Zhou, Mian [1 ]
Zhang, Yuanxing [1 ,2 ]
机构
[1] East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China
[2] Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China
[3] Roche R&D Ctr China Ltd, Shanghai, Peoples R China
基金
中国国家自然科学基金;
关键词
carbon source; Pichia pastoris; promoter; recombinant protein expression; EXPRESSION; GENE; SEQUENCE;
D O I
10.1002/yea.3301
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Pichia pastoris expression system has been widely used in recombinant protein production. So far the majority of heterologous proteins are expressed by methanol inducible promoter P-AOX1 and constitutive promoter P-GAP. The use of other promoters is rather limited. Here we selected 16 potentially efficient and regulatory promoter candidates based on the RNA-seq and RNA folding free energy G data. GFP and recombinant amylase were inserted after these promoters to reveal their strength and efficiency under different carbon sources and culture scales. Two novel promoters were successfully identified and could possibly be applied in recombinant protein expression: the methanol-inducible promoter P-0547 and the constitutive promoter P-0472.
引用
收藏
页码:379 / 385
页数:7
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