Table 1.
Plasmids and bacterial strains used in this study.
| Strain or plasmid | Relevant genotype# | References |
|---|---|---|
| E. coli | ||
| SM10 λ pir | RP4-2-Tc::Mu, π replicase (pir); strs kans | Simon et al., 1983 |
| SM10 pLP704 | pGP704:: ptlL32::kanR amp | This study |
| SM10 pBBR1L | pBBR1 MCS5::ptlL32 gen | This study |
| SM10 pCJS10 | pCJS10 cat | Dalisay et al., 2006 |
| P. tunicata D2 | ||
| WT strR | Spontaneously resistant to streptomycin | Egan et al., 2002 |
| GFP-labeled WT strR | pCJS10 cat strR | Dalisay et al., 2006 |
| ΔptlL32 | ptlL32 knockout mutant; strR, ptlL32::kanR amp | This study |
| CΔptlL32 | Complemented ptlL32 knockout mutant; str, ptlL32::kanR amp; pBBR1 MCS5ptlL32 gen | This study |
| GFP-labeled ΔptlL32 | strR, ptlL32 knockout mutant; ptlL32::kanR amp; pCJS10 cat | This study |
| PLASMIDS | ||
| pBBR1 MCS5 | Broad host range mobilizable vector, gen | Kovach et al., 1995 |
| pBBR1 MCS5::ptlL32 | Complementation vector containing intact ptlL32 | This study |
| pCJS10-GFP | RSF1010 broad host range backbone, gfpmut3, cat | Rao et al., 2005 |
| pGP704 | Suicide vector, R6K ori, mob, amp | Miller and Mekalanos, 1988 |
| pLP704 | pGP704 with ptlL32::kanR knockout fragment | This study |
| pACYC177 | Cloning vector, amp kanR | Chang and Cohen, 1978 |
#
strS, streptomycin sensitive; kanS, kanamycin sensitive; strR, streptomycin resistance; kanR, kanamycin resistance; tet, tetracyclin resistance; amp, ampicillin resistance; cat chloramphenicol resistance; gen, gentamycin resistance.