23 Jul RECOMBINANT WHITE SCELESTINE, METHOD FOR PRODUCING RECOMBINANT PROTEIN OF SCREROSTIN, METHOD OF INCREASING BONE MASS OF VERTEBRATE ANIMAL
Abstract
FIELD: biotechnology; immunobiology.SUBSTANCE: invention relates to genetic engineering, biotechnology and immunobiology. Method for producing a recombinant sclerostin protein of a vertebrate is described. Vector construct is constructed to ensure stable replication and expression of the target gene in E. coli cells. Cells of the E. coli strain expressing the sclerostin gene are grown. Sclerostin protein from cell extracts of the E. coli strain using metal-chelate affinity chromatography using a nickel-containing sorbent is purified. Then the product is washed and isolated. In order to construct the vector structure, the target sclerostin gene is first obtained and cloned. To this end, a planned oligonucleotide duplex is inserted into the target gene at the BamHI and Kpn2I sites to obtain a construct comprising a sequence encoding the fusion protein of the target sclerostin and six histidins. Method for increasing the bone mass of a vertebrate, a particular mammal and a human is also described. Twice, at an interval of 20 days, immunization with the target recombinant protein sclerostin at a dose of 0.1-100 mcg sclerostin per one kilogram body weight of the vertebrate is carried out. In this case, the target sclerostin protein of the same species of vertebrate is used, which needs to increase the density of bone tissue. Invention can be used in veterinary medicine and medicine for the treatment of osteoporosis of vertebrates and humans.EFFECT: thus, the claimed group of inventions expands the arsenal of tools used for the study and treatment of osteoporosis.3 cl, 5 ex, 2 dwg
Year: 2018
Country: RU
Doc No: 2647571