TABLE 1.
Strains and plasmids used in the study
HMK, heart muscle kinase.
| Genotype | Ref. | |
|---|---|---|
| Strains | ||
| RS 257 | MC4100 galEP3 Δrac (GJ3161) | |
| RS330 | GJ3161Δrho:: KanR , with pHYD1201 shelter plasmid, AmpR | 23 |
| RS 734 | RS257 with λRS45 lysogen carrying Plac-nutR-LacZ | |
| RS862 | MG1655 WT rac− strain | 6 |
| RS1017 | RS257 with λRS45 lysogen carrying Plac-H19BnutR-trpt′-LacZ | |
| RS1263 | MG1655 K12 WT strain | |
| RS1305 | MG1655Δrac carrying marker less Δrho (Δrho::FRT) with pHYD1201 AmpR (AI) | This study |
| RS1428 | RS257 with λRS45 lysogen carrying PRM-racR-trac-lacZYA | 6 |
| RS1430 | RS257 with λRS45 lysogen carrying PRM-racR-trac-lacZYA with marker less Δrho::FRT with pHYD1201, AmpR | 6 |
| RS1458 | RS1017, flipped out Kan cassette from RS 1017 (Plac-H19B nutR-trpt′-lacZ) | 6 |
| RS1490 | Rs734 with flipped out Kan cassette made Δrho with keio collection Δrho lysate AI (Δrho::FRT) | 6 |
| RS1514 | MC4100 carrying PRM-racR-trac-lacZYA with G146D nusG in chromosome | 6 |
| RS1523 | MC4100 carrying PRM-racR-trac-lacZYA with L158Q nusG in chromosome | 6 |
| Plasmids | ||
| pHYD1201 | 3.3-kb HindIII-SalI fragment carrying rho+ subcloned from pHYD567 into HindIII SalI sites of pAM34 (pMB1; IPTG dependent replicon, AmpR) | 23 |
| pRS22 | pTL61 T with pT7A1-H-19B nutR-TR_T1T2-lacZYA, AmpR | 20 |
| pRS96 | pET21b with WT rho cloned at NdeI/XhoI site, His-tag at C-terminal; AmpR | |
| pRS604 | T7A1-λTR1 fragment cloned at Hind III site of pRS22. | 24 |
| pRS613 | pET21b with R221C rho cloned at NdeI/XhoI site, His-tag at C-terminal; AmpR | This Study |
| pRS630 | pET21b with P235H rho cloned at NdeI/XhoI site, His-tag at C-terminal; AmpR | This study |
| pRS649 | WT rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | 11 |
| pRS695 | pHYD3011 WT nusG, AmpR | 9 |
| pRS698 | pHYD3011 N-terminal domain nusG, AmpR | 9 |
| pRS699 | pHYD3011 C-terminal domain nusG, AmpR | 9 |
| pRS700 | pHYD3011 G146D nusG, AmpR | 9 |
| pRS728 | pHYD3011 L158Q nusG, AmpR | 9 |
| pRS729 | pHYD3011 V160N nusG, AmpR | 9 |
| pRS766 | pcp20 (ts), Flp recombinase, AmpR, CamR | 25 |
| pRS900 | pET 21b with R221A rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS961 | pET 21b with WT with C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS967 | P235H rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1352 | (PRM-racR-timm-lacZYA) cloned at EcoRI-SmaI of pK8628, AmpR | 6 |
| pRS1439 | V203L rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1440 | Q193A rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1444 | M205A rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1493 | R221C rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1768 | R221A rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1492 | I168V rho with its own promoter cloned at HindIII/SacI sites of pCL1920; SpR, SmR | This study |
| pRS1532 | pET 21b with I168V rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1677 | pET 21b with R221C rho cloned at NdeI/XhoI site, HMKHis tag at C-terminal; AmpR | This study |
| pRS1678 | pET 21b with R221A rho cloned at NdeI/XhoI site, HMKHis tag at C-terminal; AmpR | This study |
| pRS1676 | pET 21b with P235H rho cloned at NdeI/XhoI site, HMKHis tag at C-terminal; AmpR | This study |
| pRS1703 | pET 21b with T276C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1704 | pET 21b with S281C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1705 | pET 21b with T286C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This Study |
| pRS1706 | pET 21b with S325C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1707 | pET 21b with P235H C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1709 | pET 21b with P235H T276C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1710 | pET 21b with P235H S281C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1711 | pET 21b with P235H T286C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1712 | pET 21b with P235H T323C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pRS1713 | pET 21b with P235H S325C C202S rho cloned at NdeI/XhoI site, His tag at C-terminal; AmpR | This study |
| pK8628 | pTL61T with Plac-H19BnutR TR1-lacZAY, AmpR | 26 |