Amann E
Dev Biol Stand. 1985;59:11-22.
A series of plasmid vectors has been constructed for the regulated, high level expression of foreign genes in E. coli. The vectors express cloned genes under the control of the tac promoter, which is a hybrid of trp and lac promoter sequences. Some of our expression vectors carry in addition to the tac promoter, the efficient lacZ ribosome binding site followed by unique cloning sites. These vectors can be used to express cloned genes directly, i.e. in an unfused, mature form. A second type of vector provides, in addition to the above regulatory elements, a translation initiation codon (ATG) for the expression of genes which have been isolated in an incomplete form (for example: cDNA). A third type of vector allow readily the construction of gene fusions to the E. coli beta-galactosidase gene, which may stabilize otherwise unstable eukaryotic proteins, and thus allows the production of high amounts of specific antigens in E. coli. With the above vectors, several eukaryotic and viral proteins, including SV40 small tumor antigen, human fibroblast interferon and herpes simplex glycoproteins have been expressed.
已构建了一系列质粒载体,用于在大肠杆菌中对外源基因进行调控的高水平表达。这些载体在tac启动子的控制下表达克隆基因,tac启动子是trp和lac启动子序列的杂交体。我们的一些表达载体除了含有tac启动子外,还带有高效的lacZ核糖体结合位点,其后是独特的克隆位点。这些载体可用于直接表达克隆基因,即表达未融合的成熟形式的基因。第二种类型的载体除了上述调控元件外,还提供一个翻译起始密码子(ATG),用于表达以不完整形式分离的基因(例如:cDNA)。第三种类型的载体便于构建与大肠杆菌β-半乳糖苷酶基因的基因融合体,这可以稳定原本不稳定的真核蛋白质,从而在大肠杆菌中大量生产特定抗原。利用上述载体,已经表达了几种真核和病毒蛋白,包括SV40小肿瘤抗原、人成纤维细胞干扰素和单纯疱疹糖蛋白。