DNA FRAGMENT, METHOD OF PRAVASTATIN SODIUM PREPARING, EXPRESSION PLASMID (VARIANTS), RECOMBINANT STRAIN (VARIANTS) Russian patent published in 1999 - IPC

FIELD: molecular biology, biotechnology, generic engineering. SUBSTANCE: DNA fragment has a size 74 base pairs, not less, in the sequence N 1. Fragment is obtained by partial digestion at direction 5'--> 3'. DNA fragment shows activity of the transcription promoter in Streptomyces and corresponds to the part of 5'-nonencoding region (1 thousand base pairs) that adjoins directly to the opened frame reading in Streptomyces carbophilus encoding cytochrome P-450_SCA-2. For producing pravastatin sodium the strain Streptomyces lividans TK-21 or Streptomyces lividans SANK 62795 transformed with gene P-450_SCA-2 expressing vector of Streptomyces carbophilus, 5'-nonencoding region (1 thousand base pairs) and DNA fragment showing the promoter activity is cultured. Culturing is carried out in the medium containing sodium-ML-236 B and under conditions providing the expression of cytochrome P-450_SCA-2. Pravastatin sodium is extracted from transformants and/or cultural medium. Expression plasmids are constructed on the base of plasmid pIJ-702 inserting DNA fragments in its showing the promoter activity. Recombinant strain Streptomyces lividans is transformed with plasmid for producing P-450_SCA-2. EFFECT: producing the useful proteins at the industrial scale. 25 cl, 5 dwg, 2 tbl, 5 ex