RECOMBINANT PLASMID DNA, METHOD OF EXPRESSION OF α-AMYLASE GENE IN STREPTOMYCES CELLS Russian patent published in 1999 - IPC

FIELD: biotechnology, molecular biology, genetic engineering. SUBSTANCE: recombinant plasmid DNA pULDVI consists of vector plasmid pIJ-699 and DNA sequence. DNA sequence is inserted into Bgl II-site of plasmid pIJ-699 and has SAF-promoter and a sequence operably bound with 5'-end of complete sequence of amy-gene located in Hind III-fragment (size is 4.8 kb) from genome DNA of S. griseus. Plasmid DNA has unique sites: for Hind III, Nco I, Bst EII, Sal I and Bgl II. Invention describes an isolation of complete sequence of gene encoding α-amylase containing promoter, site encoding signal peptide and a sequence encoding an enzyme protein. The indicated gene sequence is bound with SAF-promoter and inserted into vector followed by transformation with obtained recombinant vector of Streptomyces strain. Transformants are selected, cultured their under conditions providing expression of the inserted gene. EFFECT: improved expression of foreign DNA sequence in Streptomyces cells. 3 cl, 3 tbl, 15 dwg