Production and purification of recombinant human glucagon overexpressed as intein fusion protein in Escherichia coli.
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.
Esipov
Roman S
RS
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia. esipov@ibch.ru
Stepanenko
Vasily N
VN
Gurevich
Alexandr I
AI
Chupova
Larisa A
LA
Miroshnikov
Anatoly I
AI
D000595:Amino Acid Sequence; D004586:Electrophoresis; D004926:Escherichia coli / Q000378:metabolism; D005934:Glucagon / Q000096:biosynthesis*; D006801:Humans; D047668:Inteins / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D005934:Glucagon