Table 1.

Strains and plasmids

Species or plasmidStrain or replicon(s); host rangeGenotype or markers; characteristics and usesSource or reference(s)
E. coli DH5α supE44ΔlacU16980 lacZΔM15)hsdR17 recA1 endA1 gyrA96 thi-1 relA1 5, 11
JM109 recA1 supE44 endA1 hsdR17 gyrA96 relA1 thiΔ(lac-proAB) F′[traD36 proAB+lacI q lacZΔM15]31
G. sulfurreducens DL1Wild type, derived from single colony isolated on an NBAFYE plate7
DL1/pCD354DL1 containing pCD354This work
DL2A nifD1::kan This work
DL2B nifD1::kan This work
DL2C nifD1::kan This work
DL2D nifD1::kan This work
DL2D/pCDSnifDDL2 containing pCDSnifD This work
Plasmids
 pBBR1MCS-2ND,abroadKanr; unstable in G. sulfurreducens; source of kanamycin resistance cassette forpBRnif::kan 12
 pBR322pMB1;E. coli Ampr Tetr; suicide vector in G. sulfurreducens 6
 pBRnif pMB1;E. coli Ampr; G. sulfurreducens nifH/nifD PCR fragment cloned into pBR322(ΔEcoRI-BamHI); intermediate in the construction of pBRnif::kan This work
 pBRnif::kan pMB1; E. coli Ampr Kanr; kanamycin resistance cassette cloned into EcoRV site of pBRnif; suicide vector for construction ofnifD1::kan mutants in G. sulfurreducens This work
 pCD342IncQ; broadKanr; expression vector; suitable for use inG. sulfurreducens This work
 pCD354IncQ; broadKanr; gfpmut2 expression vector; suitable for use in G. sulfurreducens This work
 pCDnifD IncQ; broadKanr; G. sulfurreducens nifD coding sequence cloned into pCD342 (ΔHindIII-EcoRI); intermediate in the construction of pCDSnifD This work
 pCDSnifD IncQ; broadStrr; streptomycin resistance cassette cloned into pCDnifDBstBI-BglII); nifD expression vector for G. sulfurreducens nifD1::kanmutantsThis work
 pBMK7pMB1, pBG1; E. coli, Desulfovibrio spp.Kanr; E. coli andDesulfovibrio sp. shuttle vector; suicide vector in G. sulfurreducens 23
  • a ND, the incompatibility group of pBBR1MCS-2 has not been defined (2).