TABLE 1.
Strains, plasmids, and transposons used in this study
| Strain, plasmid, or transposon | Relevant characteristicsa | Reference, source, and/or construction |
|---|---|---|
| Strains | ||
| S. meliloti | ||
| RCR2011 | SU47 wild type | Rothamsted Experimental Station |
| Rm1021 | SU47 str-21 | 19 |
| Rm1021 derivatives | ||
| Rm11104 | age-1::Tn5 | 8 |
| Rm11107 | bdhA1::Tn5 | 8 |
| Rm11143 | age-1::Tn5-233 | 8 |
| Rm11159 | bdhA2::Tn5 | 1 |
| Rm11172 | age-1::Tn5-Tp | This study; Tn5-Tp replacement of Tn5-233 insertion in Rm11143 |
| Rm11175 | age-1::Tn5-Tp bdhA2::Tn5 | This study; φRm11159→Rm11172 |
| Rm11192 | bdhA2::Tn5 shb-1 | This study; spontaneous growth on DLHB |
| Rm11194 | bdhA2::Tn5 shb-2 | This study; spontaneous growth on DLHB |
| Rm11196 | bdhA2::Tn5 shb-3 | This study; spontaneous growth on DLHB |
| Rm11198 | bdhA2::Tn5 shb-4 | This study; spontaneous growth on DLHB |
| Rm11218 | bdhA2::Tn5 Ω218::Tn5-233 Hbu− | φTn5-233 bank→Rm11192 |
| Rm11219 | bdhA2::Tn5 Ω219::Tn5-233 Hbu− | φTn5-233 bank→Rm11196 |
| Rm11223 | bdhA2::Tn5 Ω223::Tn5-233 Hbu− | φTn5-233 bank→Rm11198 |
| Rm11228 | bdhA2::Tn5 Ω218::Tn5-233 shb-1 Hbu+ | φRm11218→Rm11192 |
| Rm11229 | bdhA2::Tn5 Ω219::Tn5-233 shb-3 Hbu+ | φRm11219→Rm11196 |
| Rm11232 | bdhA2::Tn5 Ω223::Tn5-233 shb-4 Hbu+ | φRm11223→Rm11198 |
| Rm11281 | bdhA1::Tn5age-1::Tn5-Tp | φRm11107→Rm11172 |
| Rm11364 | acsA27::Tn5 acsA115::Tn5-233; deficient in AACS and acetyl-CoA synthetase activities | 20 |
| E. coli | ||
| NovaBlue | endA1 hsdR17 (rK-12− mK-12+) supE44 thi-1 recA1 gyrA96 relA1 lac[F′ proA+B+ lacIqZΔM15::Tn10] Tcr | Novagen |
| BL21(DE3)pLysS | F−ompT hsdSB (rB− mB−) gal dcm (DE3) pLysS (Cmr) | Novagen |
| Plasmids | ||
| pSP329 | IncP cloning vector; Tcr | 6; S. Porter |
| pGQ105 | pSP329 carrying the S. meliloti acsA2 gene on a 4.0-kb KpnI fragment | 5 |
| pET30 Xa/LIC | N-terminal His tag expression vector; Kanr | Novagen |
| pRD112 | pET30 Xa/LIC expressing His-tagged acsA2 | This study |
| Transposons | ||
| Tn5 | Nmr Smr | 3 |
| Tn5-233 | Gmr-Spr | 11 |
| Tn5-Tp | Tpr | 16; S. Klein |
a
Abbreviations for antibiotic resistance phenotypes: Cm, chloramphenicol; Gm, gentamicin; Kan, kanamycin; Nm, neomycin; Sm, streptomycin; Sp, spectinomycin; Tc, tetracycline; Tp, trimethoprim. Abbreviations for other phenotypes: Hbu+, growth on M9 containing DLHB; Hbu−, no growth on M9 containing DLHB.