Strains or plasmids | Relevant characteristics | Reference or source |
---|---|---|
Strains | ||
E. coli TOP10 | Host strain for cloning | Invitrogen |
E. coli ET12567/pUZ8002 | Methylation defective, strain used in E. coli-streptomyces intergeneric conjugation | [25] |
E. coli BL21(DE3) | Host strain for protein expression | Novagen |
M. echinospora | Wild-type strain, gentamicin C1a, C2, C2a, and C1 producer | This lab |
M. echinospora ΔgenK | M. echinospora in which genK was disrupted | This lab |
M. echinospora ΔgenB4 | M. echinospora in which genB4 was disrupted | This study |
M. echinospora ΔgenKΔgenB4 | M. echinospora in which genK and genB4 were disrupted | This study |
M. echinospora ΔgenB4::ΔgenB4 | Complementation of genB4 in M. echinospora ΔgenB4 | This study |
M. echinospora ΔgenKΔgenB4::ΔgenB4 | Complementation of genB4 in M. echinospora ΔgenKΔgenB4 | This study |
Plasmids | ||
pIJ2925 | Cloning vector for E. coli ori(pUC18), AmpR | [26] |
pD2925 | E. coli-Streptomyces shuttle vector, oriT(RP4), ori(pUC18), AmpR, AmR | [21] |
pPT2925 | pIJ2925 containing promoter PhrdB and terminator To, AmpR | This lab |
pD2925B4 | pD2925 containing upstream and downstream fragments of genB4, AmpR, AmR | This study |
pEAP1 | E. coli-Streptomyces shuttle vector, oriT(RP4), ori(pUC18), int-attP(φC31), AmpR, ErmR | This lab |
pEAP1B4 | pEAP1 containing promoter PhrdB, genB4 fragment and terminator To, AmpR, ErmR | This study |
pET28a(+) | Protein expression vector used in E. coli, encoding N-terminal His-tag, KmR | Novagen |
pET28aB4 | pET28a(+) containing genB4 fragment, KmR | This study |
pET28aB1 | pET28a(+) containing genB1 fragment, KmR | This lab |
pET28aB2 | pET28a(+) containing genB2 fragment, KmR | This lab |
pET28aB3 | pET28a(+) containing genB3 fragment, KmR | This lab |