Arthur P M, Duckworth B, Seidman M
Otsuka Pharmaceutical Co., Ltd., Rockville, Maryland 20850.
J Biotechnol. 1990 Jan;13(1):29-46. doi: 10.1016/0168-1656(90)90129-y.
A recombinant bacterial strain for the large scale production of human interleukin-1 beta (IL-1 beta) was constructed. The lambda Pr and the tryptophan systems were compared for efficiency of transcription and regulation. The efficiency of IL-1 protein production from these constructs was analyzed. Enhanced protein synthesis was achieved by the fusion of lambda Pr promoter sequences with trp leader sequences which included the trp RBS. A strain (JM101) was selected for use as a host and tested in a one liter bioreactor. A growth and induction regimen was established for use in bioreactors which results in the accumulation of 0.75-0.95 g l-1 of recombinant IL-1.
构建了一种用于大规模生产人白细胞介素-1β（IL-1β）的重组菌株。比较了λPr和色氨酸系统在转录和调控效率方面的差异。分析了这些构建体产生IL-1蛋白的效率。通过将λPr启动子序列与包含色氨酸核糖体结合位点（trp RBS）的trp前导序列融合，实现了蛋白质合成的增强。选择了一种菌株（JM101）作为宿主，并在一升生物反应器中进行了测试。建立了一种用于生物反应器的生长和诱导方案，该方案可使重组IL-1的积累量达到0.75 - 0.95 g l-1。
