Schauder B, Blöcker H, Frank R, McCarthy J E
Gene. 1987;52(2-3):279-83. doi: 10.1016/0378-1119(87)90054-0.
New expression vectors were constructed for use in strains of Escherichia coli. Their most important feature is a polylinker system that facilitates the insertion of a gene in an optimal relationship to the highly efficient E. coli atpE translational initiation region (from nucleotide -50 to the start codon). Three ATG-containing restriction endonuclease sites can be used for the insertion of the 5' end of a gene at, or near to, its translational initiation codon. These sites may alternatively be used for the creation of a suitable translational start codon. Transcription is started by the bacteriophage lambda major promoters pR and pL in tandem and terminated by the bacteriophage fd terminator. Transcriptional initiation is very effectively repressed at 28-30 degrees C by the product of the bacteriophage lambda cIts857 gene, which is also present on the vectors. Full induction is achieved by shifting the incubation temperature to 42 degrees C. The combination of highly efficient transcriptional and translational signals on these vectors allowed high-level expression of sequences encoding human interferon beta and interleukin 2 and of the E. coli atpA, sucC and sucD genes.
构建了用于大肠杆菌菌株的新型表达载体。它们最重要的特征是一个多克隆位点系统，该系统有助于将基因以与高效的大肠杆菌atpE翻译起始区域（从核苷酸-50到起始密码子）的最佳关系插入。三个含ATG的限制性内切酶位点可用于在基因的翻译起始密码子处或其附近插入基因的5'端。这些位点也可用于创建合适的翻译起始密码子。转录由噬菌体λ主要启动子pR和pL串联启动，并由噬菌体fd终止子终止。转录起始在28-30摄氏度时被噬菌体λcIts857基因的产物非常有效地抑制，该基因也存在于载体上。通过将培养温度转移到42摄氏度可实现完全诱导。这些载体上高效转录和翻译信号的组合使得编码人干扰素β和白细胞介素2的序列以及大肠杆菌atpA、sucC和sucD基因能够高水平表达。
