METHOD OF INSERTION OF THE FOREIGN GENES TO GENOMES OF GRAM-NEGATIVE MICROORGANISMS, PLASMID VECTOR PKC 47M FOR INSERTION OF FOREIGN GENES TO GENOMES OF GRAM-NEGATIVE MICROORGANISMS, METHOD OF PLASMID VECTOR PKC 47M CONSTRUCTING

摘要

FIELD: applied genetics, microbiology. SUBSTANCE: method involves the use of plasmid vector pKC 47m (size is 22.9 t.n.p.) and capacity 5.5 mDa. Vector can be inserted to the gram-negative microorganisms by transformation and mobilization with conjugating plasmids. Method involves the construction of recombinant plasmid pKC 47m and involves the following events: cloning in vitro genes A and B of phage Mu on plasmid showing capability to replication in gram-negative microorganism followed by replacement of the own termoinducible promoter of the phage Mu with the constitutive promoter of β-lactamase. Then A^-B^--phage-mini-Mu d1 5086-1 is inserted to genome in vivo. EFFECT: improved method of gene insertion. 4 cl, 2 dwg, 1 tbl