FIELD: biotechnology, microbiology, genetic engineering. SUBSTANCE: invention relates to the batch fermentation with additional feeding with special system vector-host E. coli for effective formation of recombinant proteins, especially, recombinant antigen molecules, preferably antibody fragments, foe example, miniantibodies. Under these conditions E. coli cells grow with maximal specific growth rate up to very high density cell values. Limitations of growth due to substrates or metabolic bi-side product did not occur. Recombinant vector comprises terminating sequence in proximal and distal position, system lac-promoter/operator, sequence Shine- Delgarno T7g10, signal sequence pe1B or ompA, VH- and VL-chains of murine humanized antibody Mab425. By transformation E. coli RV308 with recombinant vector the transformed strain (pHKK) for expression of foreign protein is prepared. Invention allows to prepare large amounts of recombinant proteins. EFFECT: improved method of preparing, high yield of proteins. 12 cl, 6 dwg, 1 tbl, 5 ex
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Authors
Dates
2003-03-27—Published
1996-11-28—Filed