Xu Ning, Zhu Jinxiang, Zhu Qiaoyun, Xing Yanzi, Cai Menghao, Jiang Tianyi, Zhou Mian, Zhang Yuanxing
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Roche R&D Center (China) Ltd, Pudong, Shanghai, China.
Yeast. 2018 May;35(5):379-385. doi: 10.1002/yea.3301. Epub 2018 Feb 8.
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 and constitutive promoter P . 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 ΔG 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 and the constitutive promoter P .
毕赤酵母表达系统已广泛应用于重组蛋白生产。到目前为止,大多数异源蛋白是由甲醇诱导型启动子P和组成型启动子P表达的。其他启动子的使用相当有限。在此,我们基于RNA测序和RNA折叠自由能ΔG数据选择了16个潜在高效且具有调控作用的启动子候选序列。将绿色荧光蛋白(GFP)和重组淀粉酶插入这些启动子之后,以揭示它们在不同碳源和培养规模下的强度和效率。成功鉴定出两个新型启动子,它们可能应用于重组蛋白表达:甲醇诱导型启动子P和组成型启动子P。