Edge M D, Greene A R, Heathcliffe G R, Moore V E, Faulkner N J, Camble R, Petter N N, Trueman P, Schuch W, Hennam J
Nucleic Acids Res. 1983 Sep 24;11(18):6419-35. doi: 10.1093/nar/11.18.6419.
A 511-base pair DNA fragment encoding human interferon-alpha 2 has been chemically synthesised and expressed from a lac UV5 and a synthetic trp promoter in Escherichia coli. The synthesis involved preparation of 68 oligodeoxyribonucleotides and their enzymic ligation. The product expressed from the trp promoter system had high antiviral activity and displayed biological effects similar to those of Namalwa interferon on natural killer cell activity and in a Daudi cell growth inhibition assay. E.coli minicells containing plasmid DNA with the synthetic IFN-alpha 2 gene under trp promoter control produce a protein with the same electrophoretic mobility as a sample of authentic IFN-alpha 2. The protein from E.coli cross-reacts with the monoclonal antibody NK-2 and was readily purified, close to homogeneity, by immunoadsorption chromatography on NK-2 sepharose.
编码人α-2干扰素的一段511个碱基对的DNA片段已通过化学合成，并在大肠杆菌中由lac UV5启动子和合成的色氨酸启动子表达。合成过程包括制备68个寡脱氧核糖核苷酸及其酶促连接。由色氨酸启动子系统表达的产物具有高抗病毒活性，并且在自然杀伤细胞活性和Daudi细胞生长抑制试验中显示出与Namalwa干扰素相似的生物学效应。在色氨酸启动子控制下含有合成IFN-α2基因的质粒DNA的大肠杆菌微小细胞产生一种蛋白质，其电泳迁移率与天然IFN-α2样品相同。来自大肠杆菌的这种蛋白质与单克隆抗体NK-2发生交叉反应，并且通过在NK-2琼脂糖上的免疫吸附色谱法很容易纯化至接近均一。
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