Article Figures & Data
Figures
- Fig. 1.
Growth profiles of three SS9 strains, EC10 (▴), EC1002 (▾), and KB3 (●), cultured at 9°C at 1 atm (a) or 280 atm (b).
- Fig. 2.
Micrographs of DAPI-stained cells of P. profundum SS9. All cultures were grown at 280 atm and harvested at early stationary phase (OD600 = 0.51). (a) EC10; (b) EC1002; (c) KB3. Magnification, ×1,250. Bar, 10 μm.
- Fig. 3.
Schematic of the SS9 genomic DNA insert in pB3. The percents similarity and identity of SS9 RecBD to homologues fromE. coli (14, 15), Haemophilus influenzae (16), and M. tuberculosis(34) are shown. Several open reading frames were found on pB3 that were homologous to extracellular protein secretion genes found in E. coli (17) and V. cholerae(33, 36), among others. The BglII andPstI sites delineate the region of DNA that is cloned into pKB26 and pKB27. The thin line that separates epsH andrecD represents pMUT100 (not drawn to scale).
- Fig. 4.
Stability of plasmid pKT231 in EC10 (▴) and EC1002 (▾).
- Fig. 5.
Growth profiles of two different recDdisruption mutants created in SS9, KB60 (●) and KB63 (○), compared with EC10 (▴) and EC1002 (▾), at 280 atm and 9°C.
- Fig. 6.
High-pressure growth profiles of E. colistrains. (a) MG1655 (▴), MG1655(pUC18) (▾), MG1655(pKB27) (●), and MG1655(pKB62) (○) cultured at 310 atm and 37°C. (b) CAG12135 (▴), CAG12135(pUC18) (▾), CAG12135(pKB27) (●), and CAG12135(pKB62) (○) cultured at 310 atm and 37°C.
- Fig. 7.
Micrographs of DAPI-stained cells of E. coli. Cultures were grown at 310 atm and 37°C, and cells were harvested at early stationary phase (OD600 = 0.73). (a) CAG12135(pUC18). (b) CAG12135(pKB27). Bar, 10 μm. Magnification, ×1,250.
Tables
- Table 1.
Bacterial strains and plasmids used in this study
Strain or plasmid Description Reference(s) or source Strains P. profundum SS9 EC10 RifrSS9 derivative 8 EC1002 Pressure-sensitive EC10 derivative 8 KB3 Complemented strain of EC1002, Kanr This study KB60 EC10recD disruption mutant, Kanr This study KB63 EC10 recD disruption mutant, Kanr This study E. coli DH5α recA strain used for plasmid main-tenance 19 MC1061 Host for pRK24, pRL528, and genomic library conjugal donor strain 6, 7 ED8654 Used for pRK2073 maintenance 28 MG1655 λ− 18 CAG12135 recD::Tn10 37; CGSC7429 Plasmids pRK24 Conjugal plasmid; derivative of RK2 25 pRL528 Helper plasmid, carrying mob 13 pRK2073 Carries tra genes for conjugal transfer 4 pMUT100 pBR322-derived suicide vector; Kanr 6 pCR2.1 Vector for cloning PCR products, Kanr Invitrogen pKT231 Broad-host-range vector, Kanr 3 pUC18 High-copy-number vector, Ampr 32 pGL10 Broad-host-range vector, Kanr 1a pB3 Plasmid rescue from KB3, Kanr This study pKB26 recBD fragment from pB3 in pGL10 This study pKB27 recBD fragment from pB3 in pUC18 This study pKB60 470-bp recD fragment in pMUT100 This study pKB62 recD from MG1655 in pUC18 This study pKB63 440-bp recD fragment in pMUT100 This study pKB65 recD from MG1655 in pGL10 This study pCM1 recD from EC1002 in pCR2.1 This study
