Simons G, Remaut E, Allet B, Devos R, Fiers W
Gene. 1984 Apr;28(1):55-64. doi: 10.1016/0378-1119(84)90087-8.
Several recombinant plasmids have been constructed which direct high-level synthesis of mature human interferon gamma (IFN-gamma) in Escherichia coli using the inducible leftward promoter pL of phage lambda followed by a translational initiator region derived either from the phage MS2 replicase gene or the E. coli tryptophan attenuator region. Under these conditions, IFN levels of up to 25% of the total cellular protein can be achieved. The highest levels were obtained when a terminator of transcription was cloned downstream from the IFN-gamma sequence. IFN-gamma was almost entirely found in the initial pellet fraction and not in soluble extracts. Co-induction of the lysis genes derived from phage MS2 or from phage lambda, inserted downstream from the IFN-gamma sequence, did not enhance the biological activity present in the supernatant fraction.
已经构建了几种重组质粒,这些质粒利用噬菌体λ的可诱导向左启动子pL,随后是源自噬菌体MS2复制酶基因或大肠杆菌色氨酸衰减子区域的翻译起始区域,在大肠杆菌中指导成熟人干扰素γ(IFN-γ)的高水平合成。在这些条件下,IFN水平可达总细胞蛋白的25%。当转录终止子克隆在IFN-γ序列下游时,获得了最高水平。IFN-γ几乎完全存在于初始沉淀部分,而不存在于可溶性提取物中。共诱导源自噬菌体MS2或噬菌体λ的裂解基因(插入IFN-γ序列下游),并没有增强上清液部分中存在的生物活性。