RECOMBINANT FLAGELLIN EXTRACTION AND PURIFICATION PROCESS

摘要

The invention relates to molecular biology and biotechnology and may be used to produce recombinant flagellin. Provided is a method for production of active highly-purified recombinant truncated flagellin characterized by amino acid sequence SEQ ID NO: 1, the method comprises using E.coli BL21(DE3)/pET151D-TOPO_F27 producing strain subclone which contains a gene encoding such flagellin with optimized codon composition for expression in E.coli cells, ensuring target protein yield up to 45% from the total protein, cultivating such subclone cells using 0.5 mmol IPTG target gene expression inductor, cell lysing, cell pellet precipitation, protein extraction and purification from supernatant in denaturating conditions using Ni-NTA sepharose. The process allows production of 98% pure flagellin with a yield of 800 mg per liter of culture medium. The flagellin produced by the offered process exhibits high performance in test animal model when administered both before and after irradiation.