TABLE 1.
Strains and plasmids
| Strain/plasmid | Descriptiona | Reference/source |
|---|---|---|
| Strains | ||
| P. haloplanktis TAC125 | Wild type | 1 |
| KSL2000 | rne::cat recA::Tn10 | 31 |
| AC21 | MC1061 zce-726::Tn10 | 8 |
| AC27 | MC1061 zce-726::Tn10 rne131 | 34 |
| SVK438 | KSL2000 recA+ | V. Khemici |
| SVK446 | KSL2000 recA+ Δrhlb | V. Khemici |
| S17.1 | RP4 2-Tc::Mu-Km::Tn7 pro res mod1 | 52 |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) dcm gal λ(DE3) | 53 |
| Plasmids | ||
| pAM238 | pGB2-derived plasmid | 12 |
| pAM-rne | Chromosomal PstI fragment containing the rne gene cloned into pAM238 (pSC101ori, spectinomycin resistance) | 56 |
| pVK200 | pAM238, Ec-RNase E, C-terminal FLAG | V. Khemici |
| pET15b-RneCTH | T7 expression of the RNase E CTH with an N-terminal His tag | 29 |
| pJB3 | Broad-host-range cloning vector, RK2 derivative plasmid | 2 |
| pIB3 (shuttle vector) | Fragment 1914 to 2786 of the resident plasmid pMtBL of P. haloplanktis containing the origin of replication was cloned into pJB3 | Ivona Bagdiul |
| pSAB1 | pET15b, Ph-rne CDS under the control of T7 expression signals | This work |
| pSAB11 | pAM238, lactose expression signals deleted | This work |
| pSAB12 | pSAB11, Ph-rne | This work |
| pSAB13 | pSAB11, E. coli expression signals fused to Ph-rne CDS | This work |
| pSAB15 | pSAB11, Ph-rne, C-terminal FLAG | This work |
| pSAB17 | pIB3, Ph-rne, C-terminal FLAG | This work |
| pSAB18 | pSAB15, E. coli expression signals fused to Ph-rne CDS | This work |
| pSAB29 | pSAB18, Ph-rne (deletion of HBS) | This work |
| pSAB20 | pSAB1, Ph-rne (deletion of residues 11-528) | This work |
| pSAB33 | pSAB20, Ph-rne CTH (deletion of residues 841-1071) | This work |
| pSAB34 | pSAB20, Ph-rne CTH (deletion of residues 996-1071) | This work |
| pSAB35 | pSAB20, Ph-rne CTH (deletion of residues 1035-1071) | This work |
a
CDS, coding sequence.