Table 2.
Oligonucleotides, strains, and plasmids used in this study
| Name | Sequence 5′–3′ | Purpose |
|---|---|---|
| ccmC-F.BglII | GCGAGATCTATGTGGAAAACACTGCATC | pSysI-ccmC derivatives |
| ccmE-R. NdeI/EcoRI | GCGAATTCGGCATCATATGGCTGGGTCCTTATAAAC | pSysI-ccmC derivatives |
| ccmC H60A.R | GCCGCAGGCACAGCCAGGTAG | pRGK392 [pGEXccmC (H60A/H184A)DE] |
| ccmC W119A-R | CACCAGGTGCCCGCCATCGGTTTTC | pRGK393 [pGEXccmC (W119A)DE] |
| ccmC W125A-R | GACGTGCATCCGCTACCCACCAGGTGCC | pRGK394 [pGEXccmC (W125A)DE] |
| ccmC W125A/D126A-R | CAGACGTGCAGCCGCTACCCACCAGGTGCC | pRGK395 [pGEXccmC (W125A/D126A)DE] |
| ccmC W123A/W125A/D126A-R | CAGACGTGCAGCCGCTACCGCCCAGGTGCCCCAC | pRGK396 [pGEXccmC (W123A/W125A/D126A)DE] |
| ccmC W114A/W119A/W122A-R | CGCTACCGCCGCGGTGCCCGCCATCGGTTTTCCCGCTGCAGAGCC | pRGK397 [pGEXccmC (W5A/D126A)DE] |
| ccmC Y139A-R | ACCCACAGCCAAAAACAGCAG | pRGK398 [pGEXccmC (Y139A)DE] |
| Description | References | |
| Strains | ||
| RK103 | Δccm | 17 |
| RK105 | Δccm ΔhemA | 41 |
| RK111 | Δccm-c4:6×His integrate | Unpublished |
| Plasmids | ||
| pRGK333 | pSysI | 17 |
| pRGK354 | pSysI-ΔccmAB | 36 |
| pRGK363 | pBADccmC:6×His | 36 |
| pRGK365 | pUCA10-compccmAB | 36 |
| pRGK375 | pGEXccmCDE | 16 |
| pRGK376 | pGEXccmC (H60A)DE | 16 |
| pRGK377 | pGEXccmC (H184A)DE | 16 |
| pRGK382 | pSysI-ΔccmAC | 16 |
| pRGK383 | pSysI-ΔccmABE | 16 |
| pRGK388 | pBAD-ccmF:6×HisGH | 16 |
| pRGK391 | pGEXccmC (H60A/H184A)DE | This work |
| pRGK392 | pGEXccmC (W119A)DE | This work |
| pRGK393 | pGEXccmC (W125A)DE | This work |
| pRGK394 | pGEXccmC (W125A/D126A)DE | This work |
| pRGK395 | pGEXccmC (W123A/W125A/D126A)DE | This work |
| pRGK396 | pGEXccmC (W5A/D126A)DE | This work |
| pRGK397 | pGEXccmC (Y139A)DE | This work |