Production and Purification of Recombinant Human Glucagon Overexpressed as Intein Fusion Protein in Escherichia coli
Roman S. Esipov, Vasily N. Stepanenko, Alexandr I. Gurevich, Larisa A. Chupova and Anatoly I. Miroshnikov
Affiliation: Shemyakin-OvchinnikovInstitute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997 Russia.
Keywords: Glucagon, mini-intein, autocatalitic cleavage, recombinant, polypeptide
Chemico-enzymatic synthesis and cloning in Esherichia coli of an artificial gene coding human glucagon was performed. Recombinant plasmid containing hybrid glucagons gene and intein Ssp dnaB from Synechocestis sp. was designed. Expression of the obtained hybrid gene in E. coli, properties of the formed hybrid protein, and conditions of its autocatalytic cleavage leading to glucagon formation were studied.
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