尿素
重组DNA
尿素酶
化学
诱导剂
绿色荧光蛋白
奇异变形杆菌
表达式向量
生物制药
生物化学
分子生物学
生物
基因
生物技术
大肠杆菌
作者
Joanne Hothersall,Alexander Osgerby,Rita E. Godfrey,Tim W. Overton,Stephen J. W. Busby,Douglas F. Browning
标识
DOI:10.1016/j.nbt.2022.10.003
摘要
We have developed a novel urea-inducible recombinant protein production system by exploiting the Proteus mirabilis urease ureR-ureD promoter region and the ureR AraC-family transcriptional regulator. Experiments using the expression of β-galactosidase and green fluorescent protein (GFP) showed that promoter activity is tightly regulated and that varying the concentration of urea can give up to 100-fold induction. Production of proteins of biopharmaceutical interest has been demonstrated, including human growth hormone (hGH), a single chain antibody fragment (scFv) against interleukin-1β and a potential Neisserial vaccine candidate (BamAENm). Expression levels can be fine-tuned by temperature and different urea concentrations, and can be induced with readily available garden fertilisers and even urine. As urea is an inexpensive, stable inducer, a urea-induced expression system has the potential to considerably reduce the costs of large-scale recombinant protein production.
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