Table 2 Strains and plasmids used in this study
From: Assembly of a novel biosynthetic pathway for gentamicin B production in Micromonospora echinospora
Strains or plasmids | Relevant characteristic | Reference or source |
|---|---|---|
Strains | ||
E. coli TOP10 | F-mcrAΔ(mrr-hsdRMS-mcrBC), φ80lacZΔM15,ΔlacX74, deoR, recA1, araD139Δ(ara-leu)7697, galU, galK, rpsL(StrR), endA1, nupG | Invitrogen |
E. coli ET12567/pUZ8002 | Methylation defective, strain used in E. coli-streptomyces intergeneric conjugation | [25] |
S. kanamyceticus | Kanamycin producing strain | CGMCC4.1441 |
M. echinospora | Wild-type strain, gentamicin C1a, C2, C2a, and C1 producer | ATCC 15835 |
M. echinosporaΔP | M. echinospora with disrupted genP | [7] |
M. echinosporaΔKΔP | M. echinospora with disrupted genK and genP | This study |
M. echinospora JK1 | Heterologous, genome-based expression of kanJ and kanK, with replacement of the native promoter by PermE* in M. echinosporaΔKΔP. | This study |
M. echinospora JK2 | M. echinosporaΔKΔP + heterologous expression of kanJ and kanK under the promoter PermE*, insertion at genP locus in M. echinospora ΔKΔP | This study |
M. echinospora JK3 | Heterologous expression of kanJ and kanK under promoter PgenP, insertion at genP locus in M. echinospora ΔKΔP | This study |
M. echinospora JK4 | Heterologous expression of kanJ and kanK under promoter PhrdB, insertion at genP locus in M. echinospora ΔKΔP | This study |
Plasmids | ||
pKC1139 | E. coli-streptomyces shuttle vector, AmR | [26] |
pSPU241 | pIJ2925 derivative carrying the Streptomyces constitutive promoter PermE* and To terminator, AmpR | [8] |
pEAP1 | pSET152 carrying ermE, the apramycin resistance-conferring gene aac(3)IV was replaced by the ampicillin resistance-conferring gene bla, AmpR, ErmR | [8] |
pSPU503 | pKC1139 carrying homologous arms of genK (gacD), used in genK disruption | [12] |
pJK1 | pEAP1 carrying PermE*-kanJ-kanK, used in generating M. echinospora JK1 | This study |
pJK2 | pKC1139 carrying homologous arms and kanJK, used in generating M. echinospora JK2 | This study |
pJK3 | pKC1139 carrying homologous arms and PermE*-kanJ-kanK, used in generating M. echinospora JK3 | This study |
pJK4 | pKC1139 carrying homologous arms andPhrdB-kanJ-kanK, used in generating M. echinospora JK4 | This study |