Windass J D, Newton C R, De Maeyer-Guignard J, Moore V E, Markham A F, Edge M D
Nucleic Acids Res. 1982 Nov 11;10(21):6639-57. doi: 10.1093/nar/10.21.6639.
An 82 base pair DNA fragment has been synthesised which contains the E. coli trp promoter and operator sequences and also encodes the first Shine Dalgarno sequence of the trp operon. This DNA fragment is flanked by EcoRI and ClaI/TaqI cohesive ends and is thus easy to clone, transfer between vector systems and couple to genes to drive their expression. It has been cloned into plasmid pAT153, producing a convenient trp promoter vector. We have also joined the fragment to a synthetic IFN-alpha 1 gene, using synthetic oligonucleotides to generate a completely natural, highly efficient bacterial translation initiation signal on the promoter proximal side of the IFN gene. Plasmids carrying this construction enable E. coli cells to express IFN-alpha 1 almost constitutively and with significantly higher efficiency than from a lacUV5 promoter based system.
已合成了一个82个碱基对的DNA片段，它包含大肠杆菌色氨酸启动子和操纵序列，还编码色氨酸操纵子的第一个夏因-达尔加诺序列。该DNA片段两侧是EcoRI和ClaI/TaqI粘性末端，因此易于克隆、在载体系统之间转移并与基因连接以驱动其表达。它已被克隆到质粒pAT153中，产生了一个方便的色氨酸启动子载体。我们还使用合成寡核苷酸将该片段连接到一个合成的IFN-α1基因上，在IFN基因启动子近端一侧产生一个完全天然、高效的细菌翻译起始信号。携带这种构建体的质粒能使大肠杆菌细胞几乎组成型地表达IFN-α1，且效率明显高于基于lacUV5启动子的系统。
