Plasmids of Corynebacterium glutamicum and the use thereof

摘要

The mutually compatible plasmids pTET3 and pCRY4, isolated from Corynebacterium glutamicum DSM 5816 are new. pTET3 contains about 27.8 kb; includes a replication region of 4539 bp (1) and a 7136 bp antibiotic resistance region (6) containing the tetA gene (resistance to tetracycline) and the aadA gene (resistance to streptomycin and spectinomycin. pCRY4 contains about 48 kp and includes a replication region of 1856 bp (4). Restriction maps and sequences for (1), (4) and (6) are reproduced. Independent claims are also included for the following: (a) composite plasmid (CP1) for autonomic replication in corynefrom bacteria containing at least part of the nucleic acid sequences of these plasmids; at least one replication region derived from pTET3 or pCRY4; a replicable gene fragment from Escherichia coli, Bacillus subtilis or Streptomyces, and at least one region for expression of resistance; (b) composite plasmid (CP2) for autonomic replication in corynefrom bacteria containing at least one replication region from pGA1, pGA2, pTET3 or pCRY4; at least one resistance from pTET3 and optionally at least one DNA fragment that encodes a protein involved in biosynthesis of a vitamin, nucleotide or L-amino acid and is expressible in coryneforms; (c) plasmid vectors pSELF3-1 (about 7 kb) and pSELF1-1 (about 7.3 kb) (restriction maps reproduced); (d) production of L-amino acids (particularly lysine or threonine) by culturing a coryneform strain that contains at least one of CP1, CP2 or pSELF3-1; and (e) production of vitamins, especially D-panthothenic acid, by culturing a coryneform strain that contains at least one of pSELF3-1 or pSELF1-1.