TABLE 1.
Growth of bacterial strains
| Strain/plasmid(s) | Gene producta–e | Antibiotic resistancef | Growth on succinateg | Source or reference |
|---|---|---|---|---|
| KM2/pTAM37/pTAM46 | bhis/bV5 | Ap+Cm | +++ | Ref. 22 |
| KM2/pBR322 | Δb | Ap | − | New England Biolabs |
| KM2/pSBC127 | bV5 | Ap | ++ | This study |
| KM2/pSBC128 | bI76C, V5 | Ap | ++ | This study |
| KM2/pSBC129 | bR83C, V5 | Ap | ++ | This study |
| KM2/pSBC130 | bA90C, V5 | Ap | ++ | This study |
| KM2/pSBC131 | bE97C, V5 | Ap | + | This study |
| KM2/pSBC97 | Tb | Cm | + | [23] |
| KM2/pSBC123 | TbA83C | Cm | + | This study |
| KM2/pSBC124 | TbA90C | Cm | + | This study |
| KM2/pSBC98 | Tb′ | Ap | ++ | [23] |
| KM2/pSBC125 | Tb′A83C | Ap | ++ | This study |
| KM2/pSBC126 | Tb′A90C | Ap | ++ | This study |
| KM2/pSBC97/pSBC98 | Tb/Tb′ | Ap + Cm | ++ | [23] |
| KM2/pSBC123/pSBC125 | TbA83C/Tb′A83C | Ap + Cm | ++ | This study |
| KM2/pSBC123/pSBC126 | TbA83C/Tb′A90C | Ap + Cm | ++ | This study |
| KM2/pSBC124/pSBC125 | TbA90C/Tb′A83C | Ap + Cm | + | This study |
| KM2/pSBC124/pSBC126 | TbA90C/Tb′A90C | Ap + Cm | ++ | This study |
| MM294/pSD359 | Db+GGGC | Cm | N/A | This study |
| MM294/pSD358 | Db′+GGGC | Ap | N/A | This study |
| MM294/pSD361 | PbA83C | Ap | N/A | This study |
| MM294/pSD363 | PbA90C | Ap | N/A | This study |
| MM294/pSD362 | Pb′A83C | Ap | N/A | This study |
| MM294/pSD364 | Pb′A90C | Ap | N/A | This study |
a Tb, wild-type E. coli b subunit with sequence from the b subunit of T. elongatus substituted for amino acids Glu39–Ile86, a C21S mutation, and a six-histidine epitope tag at the amino terminus.
b Tb′, wild-type E. coli b subunit with sequence from the b′ subunit of T. elongatus substituted for amino acids Glu39–Ile86, a C21S mutation, and a V5 epitope tag (GKPIPNPLLGLDST) at the carboxyl terminus.
c The native cysteine at bC21has been mutated to serine in all pSBC plasmids.
d The Db and Db′ genes produce peptides modeling the dimerization domains of the Tb and Tb′ subunits. These peptides have the sequence MSY at the amino terminus, contain Tb/Tb′ residues Asp53–Lys122in the middle, and terminate with the sequence GGGC at the carboxyl terminus.
e The Pb and Pb′ genes produce peptides modeling the D53-L156 regions of the Tb and Tb′ subunits. These peptides also start with the sequence MSY and have cysteine residues substituted at either Ala83 or Ala90 as specified.
f Ap, ampicillin; Cm, chloramphenicol.
g +++, wild-type growth; ++, colonies smaller than wild-type; +, small colony formation; −, no growth.