METHOD FOR PRODUCTION OF PROTEIN PREPARATION BELONGING TO STRESS GENUS AND HSP70 PROTEIN PREPARATION OBTAINED BY SAID METHOD Russian patent published in 2006 - IPC A61K38/12 C12N1/00 

FIELD: molecular biopharmacology.

SUBSTANCE: method for production of stress protein (heat shock protein) preparations includes such protein gene expression in E.coli cells followed by isolation, refolding and purification. Gene expression is carried out in structure of vector cDNA containing T7 promoter, kanamycin resistance gene, replicative pUC ori origin, and gene encoding lac-inductor. Vector DNA is cultured in structure of E.coli cells BL21 in nutrient medium with addition of kanamycin (100 mug/ml) and glucose (0.2 %) up to turbidity level of 0.5-0.6 OD600. Then 0.02mM isopropyl beta-D-thiogalactoside is added into medium and culturing is carried out for 1-3 h to accumulate finish product. Target product is purified by anion exchange chromatography. In said method vector DNA encoding proteins: HSP 100-200; HSP 100; HSP 90; Lon; HSP 70; HSP 60; TF 55; HSP 40; FKBPl cyclophylin; HSP 2-30; ClpP; GrpE; HSP 10, etc. of any gene are used. Invention also related to stress protein (heat shock protein) preparation obtained by abovementioned method. Protein represents recombinant HSP protein having molecular weight of about 70 kDa, which is isolated after incubation of E.coli cells in growth medium LB for 12 h at 37°C with addition of kanamycin (100 mug/ml) and glucose (0.2 %) up to turbidity level of 0.5-0.6 OD600, wherein transformed cell contains pBSH2-HSP70 plasmide DNA, carrying cDNA of heat shock protein gene, whish has size of 5118 n.p. and represents pBSH2 promoter containing T7 promoter, kanamycin resistance gene, replicative pUC ori origin, and gene encoding lac-inductor. Finished protein product has purity of at least 98 %.

EFFECT: stress proteins of high activity and high purity.

8 cl, 5 ex, 5 dwg