Remaut E, Stanssens P, Fiers W
Nucleic Acids Res. 1983 Jul 25;11(14):4677-88. doi: 10.1093/nar/11.14.4677.
We have obtained high level synthesis in Escherichia coli of mature human fibroblast interferon using a plasmid vector that was designed to allow easy coupling of a DNA coding region to the initiator AUG of the replicase gene of the RNA phage MS2 cloned downstream of phage lambda's leftward promoter. The activity of the promoter can be regulated by temperature. Induced cells accumulated the interferon up to 4% of the total cellular protein. The biological activity of the product amounted to 4 X 10(9) international units per litre of culture. The synthesis of human fibroblast interferon was shown to drastically inhibit the growth rate of the bacterial host.
我们利用一种质粒载体在大肠杆菌中实现了成熟人成纤维细胞干扰素的高水平合成。该质粒载体的设计使得编码区DNA能够轻松地与克隆在噬菌体λ左向启动子下游的RNA噬菌体MS2复制酶基因的起始密码子AUG相连。启动子的活性可通过温度调节。诱导细胞中积累的干扰素高达细胞总蛋白的4%。产物的生物活性达到每升培养物4×10⁹国际单位。结果表明，人成纤维细胞干扰素的合成会显著抑制细菌宿主的生长速率。
