TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristic(s)a | Source or reference |
|---|---|---|
| Paenarthrobacter sp. strain YJN-5 | Degrades iprodione, Strr | This study |
| Microbacterium | ||
| YJN-G | Degrades iprodione | 12 |
| YJN-5M | ipaH insertion mutant of Paenarthrobacter sp. strain YJN-5, Strr, Gmr | This study |
| YJN-5M(pBBR1-ipaH) | ipaH gene complemented by pBBR1-ipaH in YJN-5M, Strr, Gmr, Kmr | This study |
| E. coli | ||
| DH5α | F− recA1 endA1 thi-1 supE44 relA1 deoR Δ(lacZYA-argF)U169 ϕ80dlacZΔM15 | Vazyme |
| BL21(DE3) | F− ompT hsdSB(rB− mB−) dcm gal λ(DE3) | Vazyme |
| Plasmids | ||
| pMD19-T | TA clone vector, Ampr | TaKaRa |
| pUC118 BamHI/BAP | DNA library construction vector, Ampr | TaKaRa |
| pR1 | pUC118 derivative carrying 5,716-bp insertion including ipaH gene, Ampr | This study |
| pR2 | pUC118 derivative carrying 7,828-bp insertion including ipaH gene, Ampr | This study |
| pET-29a(+) | Expression vector, Kmr | Laboratory stock |
| pET-ipaH | pET-29a(+) derivative carrying ipaH, Kmr | This study |
| pEX18Gm | Gene knockout vector, oriT, sacB, Gmr | 42 |
| pEX-ipaH | ipaH gene knockout vector containing partial homologous regions of ipaH, Gmr | This study |
| pBBR1MCS-2 | Broad-host-range vector, Kmr | 44 |
| pBBR1-ipaH | ipaH gene complementation vector containing ipaH, Kmr | This study |
| pET-K82A | pET-29a(+) derivative carrying ipaH-K82A, Kmr | This study |
| pET-S157A | pET-29a(+) derivative carrying ipaH-S157A, Kmr | This study |
| pET-S181A | pET-29a(+) derivative carrying ipaH-S181A, Kmr | This study |
a
Strr, streptomycin resistant; Gmr, gentamicin resistant; Ampr, ampicillin resistant; Kmr, kanamycin resistant; Cmr, chloramphenicol resistant.