METHOD OF HIGH-LEVEL EXPRESSION, METHOD OF PURIFICATION AND METHOD OF OVER-STACKING OF PORINE PROTEIN OF MENINGOCOCCUS OF GROUP B EXTERNAL MEMBRANE OR PROTEIN FUSED WITH ITS, HIGHLY PURIFIED PORINE PROTEIN (VARIANTS), HIGHLY PURIFIED FUSED PROTEIN (VARIANTS), VACCINE FOR PROPHYLAXIS OF BACTERIAL MENINGITIDIS OF GROUP B, METHOD OF PREPARING CONJUGATE OF MENINGOCOCCUS OF GROUP B PROTEIN AND POLYSACCHARIDE, METHOD OF PROPHYLAXIS OF BACTERIAL MENINGITIDIS, STRAIN OF ESCHERICHIA COLI (VARIANTS) Russian patent published in 2002 - IPC

FIELD: invention relates to genetic engineering, molecular biology, medicine, microbiology. SUBSTANCE: method involves transformation of vector into the strain of E. coli. Vector involves a selective marker and gene encoding the matured porine protein or matured porine protein fused with amino acids 1-22 of capsid protein ⊘ 10. The latter is encodes by gene T7 that is associated with promoter T7 functionally. The transformed strain of E. coli is cultured in medium and expression of indicated protein is induced. The mass of expressed protein is 2% of total expressed proteins. Then E. coli cells are subjected for lysis at stage of inducing. Protein isolated as insoluble inclusions is washed out with buffer, suspended and inclusions are dissolved in an aqueous solution of denaturating agent, diluted with detergent and purified by gel-filtration and ion-exchange chromatography. The highly purified porine protein and fused protein show amino acid sequence given in description and they encode DNA with sequence given in description. Vaccine has porine protein of external membrane or protein fused with its taken in effective amount and pharmaceutically acceptable carrier. The prepared over-stacking protein is conjugated with polysaccharide from Neisseria meningitides followed by administration of latter in porine protein of meningococcus of group B protein or protein fused with it taken in effective amount. The strain E. coli BL21(DE3) Δ ompA is designated for preparing recombinant strain-producers of porine protein of meningococcus of group B or protein fused with its. After transformation of this strain with plasmid vector pET carrying gene encoding porine protein and protein fused with porine it produces porine protein and protein fused with it. Invention provides to carry out the isolation of porine proteins of external membrane Meningococcus of group B and associated proteins. EFFECT: improved method of expression and preparing. 24 cl, 11 dwg, 1 tbl