Construction of a stable bioengineered strain of biotechmycin
Shang, G.; Dai, J.; Wang, Y.
Chinese Journal of Biotechnology 15(2): 105-111
1999
ISSN/ISBN: 1042-749X PMID: 10719630 Document Number: 504433
A stable bioengineered strain of Biotechmycin (Streptomyces spiramyceticus WSJ-1) was constructed by integrating the 4"-isovaleryltransferase gene (ist) through homologous recombination into the chromosome of spiramycin-producing strain S.spiramyceticus F21. In this construction, a Streptomyces/E.coli shuttle plasmid pKC1139 (AmR) was used as the vector and tsr gene was inserted as the marker for selection of homologous recombination. This constructed strain, S.spiramyceticus WSJ-1, was genetically very stable in production titer and in the production of biotechmycin as well as in carrying tsr selective marker when grown without pressure. The fermentation of S.spiramyceticus WSJ-1 was also improved compared with the original strain harboring unintegrated plasmid. Southern hybridization confirmed the integrated status of the ist gene in the host genome.