title: Direct formation of human interleukin-11 by cis-acting system of plant virus protease in Escherichia coli.
abstract: To produce a large amount of recombinant proteins in Escherichia coli, we constructed a unique cis-acting expression system using a plant virus protease. This new expression system could directly produce recombinant proteins, that had a biologically active form. A gene of nuclear inclusion-a (NIa), which had a specific amino acid sequence, was fused with a foreign protein gene at the same protein reading frame. One of the NIa-specific cleavage amino acid sequences, Gln-Ala, was also contained at the protein-protein junction. In the case of human interleukin-11 (hIL-11), a 23-kDa specific signal band was obtained from recombinant bacteria. N-terminal sequencing of the 23-kDa protein showed that NIa specifically cleaved the fusion protein at Gln-Ala, producing Ala-hIL-11. Furthermore, we could produce the mature rhIL-11 by extending the culture time. This 23-kDa protein had the same biological activity as hIL-11 in a mouse plasmacytoma, T1165. Combined with fermentation control, we produced mature rhIL-11 in E. coli.
authors.affiliation: Biomedical Research Laboratories, Sankyo Co. Ltd., Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
authors.forename: M
authors.initials: M
authors.lastname: Nakanishi
authors.forename: Y
authors.initials: Y
authors.lastname: Yao
authors.forename: I
authors.initials: I
authors.lastname: Uyeda
authors.forename: N
authors.initials: N
authors.lastname: Serizawa
mesh_terms: D017931:DNA Primers; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D007887:Fabaceae / Q000821:virology*; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006801:Humans; D017370:Interleukin-11 / Q000235:genetics*; D010942:Plant Viruses / Q000201:enzymology*; D010946:Plants, Medicinal
chemical_list: D017931:DNA Primers; D017370:Interleukin-11; D010450:Endopeptidases