Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionSource or reference
E. colistrains
 DH5α supE44 ΔlacU169 (φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 36
 S17-1 thi pro hsdR hsdM+ recA, chromosomally integrated RP4-2 (Tc::Mu Km::Tn7) 41
Methylotroph strains
Methylobacterium sp. strain DM4(DSM 6343)DCM+ 17
Methylobacterium sp. strain DM4-2crSmrDCM, mutant of DM4 18
Methylobacterium sp. strain DM4-2cr(DM4)Smr Kmr,Methylobacterium sp. strain DM4-2cr carrying pME1683This study
Methylobacterium sp. strain DM4-2cr(DM11)Smr Kmr,Methylobacterium sp. strain DM4-2cr carrying pME1685This study
Methylophilus sp. strain DM11(DSM 6813)DCM+ 39
Plasmids
 pBluescript II KS(+)Cloning vector, Apr Stratagene
 pME15404.40 kb, 1.5-kbHindIII-PstI fragment with dcmA ofMethylobacterium sp. strain DM4 in pBluescript II KS(+) (KpnI, Bsp120I, XhoI, SalI, and ClaI missing upstream of HindIII in the polylinker) 37
 pME19194.0 kb, pET-derived expression vector with 0.9-kb DNA fragment containing thedcmA gene of strain DM11, Apr 47
 pME16733.1 kb,BamHI-XbaI trpA terminator region from the pBAce derivative pGST3a in pBluescript II KS(+) This study
 pME1671pBluescript II KS(+) derivative containing a 1,060-bp fragment with the dcmA gene ofMethylophilus sp. strain DM11, the promoterPA , and the trpA terminator as aHindIII-XbaI insertThis study
 pME1681pBluescript II KS(+) derivative containing the 1,513-bp HindIII BamHI fragment of pME1540 with the dcmA gene of Methylobacterium sp. strain DM4This study
 pJB3Km16.05 kb, broad-host-range cloning vector derived from RK2, KmrApr 4
 pME16831,557-bpKpnI-XbaI fragment of pME1681 inKpnI-XbaI fragment of pJB3Km1This study
 pME16851,115-bp KpnI-XbaI fragment of pME1671 in KpnI-XbaI fragment of pJB3Km1This study
  • a See references 9 and29.