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Human IL-36RA production in Escherichia coli with coexpression of E. coli methionine aminopeptidase. I. Comparison of IL-36RA production in different strains

A. A. Kolobov, E. V. Kondratyeva,T. V. Kudling, M. M. Karasev,R. S. Kalinin, A. A. Khizhina,P. P. Nimiritsky,V. E. Stefanov,A. V. Petrov

Cell and Tissue Biology(2017)

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Abstract
Generalized pustular psoriasis (GPP) is a rare, sometimes lethal, form of psoriasis caused by series of mutations in the interleukin-36 receptor antagonist (IL-36RA) gene associated with its reduced expression or activity. Administration of exogenous IL-36RA can be a potent therapeutic approach to treating GPP and other forms of psoriasis. Since cleavage of the starting N-formylmethionine residue from the N-terminal end is needed for full biological activity of IL-36RA, we have developed a technique for producing IL-36RA lacking N-formylmethionine residue in E. coli . We have created a series of plasmids carrying the E. coli methionine aminopeptidase (MAP) gene under the control of different promoters for coexpression of IL-36RA and MAP and tested their effect on IL-36RA production. The highest production of IL-36RA with <3% of unprocessed molecules with uncleaved N-terminal formylmethionine residue has been shown for E. coli strain carrying the MAP gene under the control of arabinose-inducible promoter.
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Key words
Interleukin, interleukin-36 receptor antagonist (IL-36RA), methionine aminopeptidase (MAP), recombinant protein, coexpression, methionine
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