Abstract
The CRM197 is a non-toxic mutant of diphtheria toxin having a single amino acid substitution of a glycine for a glutamic acid in position 52. Being naturally nontoxic, CRM197 is a promising adjuvant and ideal carrier protein for conjugate vaccines. Typically, production of diphtheria toxin and some of the non-toxic proteins are carried out by Corynebacterium diphtheriae. Production of recombinant protein CRM197 in Escherichia coli is advantageous. It is simple, cheap and permits production of the target protein in a short time using a non-pathogenic microorganism. In this study patented high-yield-producing E. coli strain was used. As a part of the study the following steps were taken: protocol adjustment for induction of crm197 gene, production and purification of recombinant CRM197 by ion-exchange, hydrophobic and gel-filtration chromatography. The purity of the final preparation reached 97%.
Highlights
The CRM197 is a non-toxic mutant of diphtheria toxin having a single amino acid substitution of a glycine for a glutamic acid in position 52.
Получение рекомбинантного белка CRM197 в клетках E. coli // Инфекция и иммунитет.
При этом необходимо выращивать культуру клеток в течение 36–48 ч для достижения максимальной концентрации дифтерийного токсина или его производных.
Summary
The CRM197 is a non-toxic mutant of diphtheria toxin having a single amino acid substitution of a glycine for a glutamic acid in position 52. Получение рекомбинантного белка CRM197 в клетках E. coli // Инфекция и иммунитет. При этом необходимо выращивать культуру клеток в течение 36–48 ч для достижения максимальной концентрации дифтерийного токсина или его производных.
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