TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| S17.1 | RP4-2 Tc::Mu Km::Tn7 | 50 |
| BL21(DE3) | Host for overexpression of GlnBHis6 and GlnKHis6 | Novagen |
| R. capsulatus | ||
| B10S | Wild-type Smr | 32 |
| BSRUB13 | ΔglnK | This work |
| PHU332 | glnB::Kmr | P. Hübner, Basel, Switzerland |
| RCAY22 | amtY::Gmr | 62 |
| RCAY63 | amtB::Kmr | 62 |
| SG21 | glnK::Gmr | This work |
| TR3 | glnB51 orf3528::Gmr | This work |
| Plasmids | ||
| pAP2 | pJB3TC20 derivative carrying glnK51 (GlnK-Y51F) and 300 bp of upstream sequence | This work |
| pAY98 | pMECA derivative carrying glnKamtB; Apr | A. Yakunin |
| pBSRUB13 | pNIRUB145 derivative carrying lacZc expressed from a Km cassette; Gmr | This work |
| pBSRUB19 | pPHU231 derivative carrying amtB and most of glnK | This work |
| pBSRUB21-I | pBSRUB19 derivative carrying amtB expressed from a Km cassette | This work |
| pET-22b(+) | T7 expression vector; Apr | Novagen |
| pJB3TC20 | Broad-host-range vector; mob Apr Tcr | 5 |
| pK18 | Cloning vector; Kmr | 46 |
| pML5 | Broad-host-range lacZ-fusion vector; mob Tcr | 35 |
| pNIRUB60-I | pUC18 derivative carrying aphII (Kmr) of transposon Tn5 | This work |
| pNIRUB142 | pML5 derivative carrying lacZc expressed from a Km cassette | This work |
| pNIRUB143 | pSUP202 derivative carrying most of amtB | This work |
| pNIRUB144 | pNIRUB143 derivative carrying glnK::GmramtB | This work |
| pNIRUB145 | pNIRUB144 derivative with 228-bp deletion in glnK | This work |
| pPHU231 | Broad-host-range vector; mob Tcr | 47 |
| pSG5IIa | pUC18 derivative carrying glnK::Gmr; Apr | This work |
| pSG18 | pK18 derivative carrying most of amtB | This work |
| pSGNJ1 | pSVB10 derivative carrying glnK-amtB; Apr | This work |
| pSUP202 | Mobilizable cloning vector; mob Tcr Apr Cmr | 50 |
| pSV5 | pET-22b(+) derivative carrying glnBHis6 | This work |
| pSV6 | pET-22b(+) derivative carrying glnKHis6 | This work |
| pSVB10 | Cloning vector; Apr | 2 |
| pTR9 | pK18 derivative carrying 5′ part of glnB and orf3528 | This work |
| pTR10 | pUC18 derivative carrying 3′ part of glnB | This work |
| pTR11 | pTR9 derivative carrying Gmr from pWKR440 inserted into orf3528 | This work |
| pTR12 | pTR10 derivative; mob Tcr | This work |
| pTR14-I | pTR12 derivative carrying glnB51 (GlnB-Y51F) and orf3528::Gmr | This work |
| pUC18 | Cloning vector; Apr | 55 |
| pWKR56-I | Mobilizable cloning vector; mob Tcr | 32 |
| pWKR440 | pACYC derivative carrying Gmr Tcr | 11 |