TABLE 2.
Plasmids used and constructed for this work
| Plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| pA1 | pK18mob carrying an SsII-KpnI insert containing the complete greA gene and the 5′ end of the lpsB gene of S. meliloti 2011; coding sequences of the insert in sense orientation relative to the plac promoter from the vector | This work |
| pA2 | pK19mob carrying the same insert as in pA1; coding sequences of the insert in antisense orientation relative to plac promoter from the vector | This work |
| pAB2001 | Plasmid containing a promoterless lacZ-aacC1 cassette conferring Gmr | Becker et al. (5) |
| pAL100 | pACYC184-Gm-mob carrying in DraI a 5.5-kb SstI DNA fragment from S. meliloti 2011 that complements the LPS mutations in S. meliloti 6963 (blunt-end cloned as Ecl136II), Gmr | Niechaus et al. (41) |
| pAL101 | pSVB30/ΔB-H carrying in SstI the 5.5-kb SstI fragment from S. meliloti 2011 that complements S. meliloti 6963 | This work |
| pAL103A | pSVB30 carrying in SalI (Klenow) the 5.5-kb SstI fragment from pAL101 blunt-end cloned as Ecl136II; insert orientation with its HindIII site close to the lac promoter from the vector | This work |
| pAL103B | pSVB30 carrying in the SalI restriction site (Klenow) the 5.5-kb SstI fragment from pAL101 blunt-end cloned as Ecl136II; insert orientation opposite that in pAL103A | This work |
| pB1 | pK18mob carrying a BamHI-KpnI insert that contains the 3′ end of the greA gene and the 5′ portion of the lpsB gene of S. meliloti 2011; coding sequences of the insert in sense orientation relative to the lac promoter from the vector | This work |
| pB2 | pK19mob carrying the same insert as in pB1; coding sequences of the insert in antisense orientation relative to the lac promoter from the vector | This work |
| pC1 | pK18mob carrying an internal fragment of the lpsB gene that extends from a few codons downstream of the putative GTG start codon to the central KpnI site; coding sequences of the insert in sense orientation relative to the lac promoter from the vector | This work |
| pC2 | pK19mob carrying the same insert as in pC1; coding sequences of the insert in antisense orientation relative to the lac promoter from the vector | This work |
| pJB3Tc19 | Derivative of the minimal replicon of plasmid RK2, Apr Tcr | Blatny et al. (8) |
| pJBIpsSme | pJB3Tc19 carrying a 4.9-kb SstI-HindIII fragment containing the greA-lpsD region from S. meliloti 2011 cloned in the polylinker of the vector as EcoRI-HindIII, Kmr | This work |
| pJL200 | pSVB30 carrying in SstI an 11-kb SstI Tn5-containing fragment from S. meliloti 6963; insert homologous to that of pAL101 but carrying the Tn5 within lpsB | Niehaus et al. (41) |
| pJL200/ΔB-H | pSVB30/ΔB-H carrying in SstI the 11-kb SstI Tn5-containing fragment from S. meliloti 6963 | This work |
| pJL201 | pACYC184-Gm-mob carrying in DraI an 11-kb SstI Tn5-containing fragment from pJL200 blunt-end cloned as Ecl136II, Nmr Gmr; insert homologous to that of pAL100 but carrying the Tn5 within lpsB | Niehaus et al. (41) |
| pK18mob | pK18 derivative, mob, Kmr | Schäfer et al. (52) |
| pK19mob | pK19 derivative, mob, Kmr | Schäfer et al. (52) |
| pPN120 | pLAFR1 with a 4.4-kb EcoRI fragment containing greA, lpcC, dctA, and the 5′ region of dctB from R. leguminosarum bv. viciae, Tcr | C. Ronson (28) |
| pSVB30 | pUC8 derivative, Apr | Arnold and Pühler (1) |
| pSVB30/ΔB-H | pSVB30 with a deletion in the BamHI-HindIII region of the polylinker | This work |