Table 1.
Microbial strains and plasmids used in this study
| Strain or plasmid | Relevant features | Source or reference |
|---|---|---|
| E. coli strains | ||
| GC5 | F- Φ80lacZM15 (lacZYA-argF)U169 recA1 endA1 hsdR1 7(rK mK+) phoA supE44 thi-1 gyrA96 relA1 tonA |
Genesee |
| BL21(DE3) | FompT hsdSB(rB mB) gal dcm (DE3) | Novagen |
| L. pneumophila strains | ||
| Lp02 | Philadelphia-1, serogroup 1, salt sensitive, restriction deficient, thymidine auxotroph; Smr |
(24) |
| Lp03 | Philadelphia-1, serogroup 1, salt sensitive, restriction deficient, thymidine auxotroph, lacking dotA3; Smr |
(27) |
| Plasmids | ||
| pCPDnHalo |
E.coli cell-free expression vector; encodes HaloTag alone; Ampr |
This study |
| pJFT6_nHalo | Wheat germ cell-free expression vector for N-temrinal HaloTag fusion; Ampr |
This study |
| pJFT7_nHalo | Mammalian cell-free expression vector for N-terminal HaloTag fusion; Ampr |
(28) |
| pJFT7_cHalo | Mammalian cell-free expression vector for C-terminal HaloTag fusion; Ampr |
(28) |
| pANT7_cGST | T7-based mammalian expression vector; encodes C-terminal GST tag; Ampr |
(29) |
| pLDNT7_nFLAG | T7-based mammalian expression vector; encodes N-terminal FLAG tag; Ampr |
(30) |
| pFN22k–lidA | Encodes LidA with N-terminal HaloTag; Kanr | This study |
| pFC15k–lidA | Encodes LidA with C-terminal HaloTag; Kanr | This study |
| pFN22k–sidM | Encodes SidM with N-terminal HaloTag; Kanr | This study |
| pFC15k–sidM | Encodes SidM with C-terminal HaloTag; Kanr | This study |
| pGEX-6p-1-sidM | Encodes SidM with N-terminal GST tag; Ampr | (7) |
| pGEX-6p1-sidMD110/ 112A | Encodes SidMD110/112A with N-terminal GST tag; Ampr | This study |
| pDEST17-lidA | Encodes His6-LidA; Ampr | This study |
| pDEST17-sidM | Encodes His6-SidM; Ampr | This study |
| pcDNA6.2/N-EmGFP- DEST |
Encodes GFP; Camr, Ampr | Life Technologies |
| pcDNA6.2/N-EmGFP- DEST-rab1B |
Encodes GFP- Rab1B (human); Ampr | This study |
| pcDNA6.2/N-EmGFP- DEST-rab8B |
Encodes GFP- Rab8B (human); Ampr | This study |
| pcDNA6.2/N-EmGFP- DEST-rab10 |
Encodes GFP- Rab10 (human); Ampr | This study |
| pcDNA6.2/N-EmGFP- DEST-rab27A |
Encodes GFP- Rab27A (human); Ampr | This study |
| pcDNA6.2/N-EmGFP- DEST-ocel1 |
Encodes GFP- OCEL1 (human); Ampr | This study |
| pmCherry-C1 | Encodes mCherry; Kanr | Clontech |
| pmCherry-lidA | Encodes mCherry-LidA; Kanr | This study |
| pmCherry-sidMD110/112A | Encodes mCherry-SidMD110/112A; Kanr | This study |
Abbreviations: Smr, streptomycin resistance; Kanr, kanamycin resistance; Ampr, ampicillin resistance; Camr, chloramphenicol resistance.