TABLE 1.

Strains and plasmids used in this studya

Strain or plasmidDescriptionbReference or source
R. eutropha strains
    Ae H16Wild-type; Gm resistantATCC17699
    Re1097ΔphaZ1 strain, derived from Ae H16/pGY96This study
    Re1107ΔphaZ3 strain, derived from Ae H16/pPK51This study
    Re1108ΔphaZ1ΔphaZ3, strain, derived from Re1097/pPK51This study
    Re1110ΔphaZ2 strain, derived from AeH16/pJOE39This study
    Re1111ΔphaZ1ΔphaZ2ΔphaZ3 strain, derived from Re1108/pJOE39This study
    Re1112ΔphaZ1ΔphaZ2 strain, derived from Re1097/pJOE39This study
    Re1113ΔphaZ2ΔphaZ3 strain, derived from Re1107/pJOE39This study
E. coli strains
    DH5αStrain for ligation, cloning, and heterologous expression of PHA genesNew England Biolabs
    DH5αF′F′/endA1 hsdR17 glnV44 thi-1 recA1 gyrA relA1DU169 deoR19
    S17-1recA pro hsdR RP4-2-Tc::Mu-Km::Tn7ATCC47055
Plasmids
    PBluescriptIIKSCloning vector; LacZa Ap resistanceStratagene
    pCR2.1-TOPOHigh-copy-number plasmid used for cloning; confers Ap and Km resistanceInvitrogen
    pJQ200mp 18KmDerivative of pJQ200mp 18; Gm resistance gene disrupted; confers Km resistance21
    pGY96ΔphaZ1 gene replacement plasmid; confers Km resistanceThis study
    pJOE39ΔphaZ2 gene replacement plasmid; confers Km resistanceThis study
    pPK51ΔphaZ3 gene replacement plasmid; confers Km resistanceThis study
    pWt-250-2∼240-bp PCR product corresponding to phaZ1; cloned in pCR2.1-TOPOThis study
    pZ15∼240-bp PCR product corresponding to phaZ2; cloned in pCR2.1-TOPOThis study
    pZ31∼240-bp PCR product corresponding to phaZ3; cloned in pCR2.1-TOPOThis study
  • a Plasmids constructed in this study that were used only as intermediates for construction of other plasmids are described only in Materials and Methods.

  • b Abbreviations: Ap, ampicillin; Gm, gentamicin; Km, kanamycin.