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• W2031141647 abstract "We describe the construction, expression characteristics and some applications of a versatile dual-promoter expression plasmid for heterologous gene expression in Escherichia coli which contains both λ PL and PT7 promoters. Furthermore, the plasmid is optimized to allow the expression of mature coding sequences without compromising the strength of the highly efficient PT7 or of the T7g10 ribosome-binding site. The effect of the the naturally occurring RNA loops at both the 5′ and 3′ ends of the T7g10 mRNA on expression was also examined. A double T7 RNA polymerase transcription terminator was inserted to ensure more reliable transcription termination and a higher expression level of the preceding gene. Further improvements involve a clockwise orientation of the promoters to minimize read-through transcription from plasmid promoters, a largely extended multiple cloning site, an antisense phage T3 promoter and a phage f1-derived, single-stranded replication origin. Variants of this plasmid allow for the production of fusion proteins with part of T7g10, a hexahistidine peptide and an enterokinase recognition site. The potential of these expression vectors is demonstrated by comparing the expression levels of a number of mammalian cytokines (human tumor necrosis factor, human immune interferon, human and murine interleukins 2, murine interleukin 4 and murine fibroblast interferon), using these expression plasmids." @default.
• W2031141647 created "2016-06-24" @default.
• W2031141647 creator A5057399836 @default.
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• W2031141647 date "1995-10-01" @default.
• W2031141647 modified "2023-10-14" @default.
• W2031141647 title "Versatile, multi-featured plasmids for high-level expression of heterologous genes in Escherichia coli: overproduction of human and murine cytokines" @default.
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• W2031141647 doi "https://doi.org/10.1016/0378-1119(95)00505-z" @default.
• W2031141647 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/7590329" @default.
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