Skip to main content

Advertisement

Table 2 Strains and plasmids used in this study

From: Modification of acetoacetyl-CoA reduction step in Ralstonia eutropha for biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from structurally unrelated compounds

Strain or plasmid Relevant marker Source or references
Escherichia coli
DH5α F, ϕ80lacZ∆M15, ∆(lacZYA-argF) U169, deoR, recA1, endA1, hsdR17(r K m K + ), phoA, supE44, λ, thi-1, gyrA96, relA1 Lab stock
BL21(DE3) E. coli B, F, dcm, ompT, hsdS(r B m B ), gal, λ(DE3) Novagen
S17-1 thi pro hsdR recA chromosomal RP4; Tra+; Tmpr Str/Spcr [43]
Ralstonia eutropha
H16 Wild type DSM 428
NSDG H16 derivative; ΔphaC::phaCNSDG [33]
NSDG-GG NSDG derivative; ΔnagR, nagE(G793C), PA2858-glpFKEc-h16_A2858 This study
NSDG-GGΔB1 NSDG-GG derivative; ΔphaB1 This study
NSDG-GG-BNV NSDG-GG∆B1 derivative; phaB1NV This study
NSDG-GG-BNVYF NSDG-GG∆B1 derivative; phaB1NVYF This study
NSDG-GG-B2 NSDG-GG∆B1 derivative; ∆phaB1::phaB2 This study
NSDG-GG-HC NSDG-GG∆B1 derivative; ∆phaB1::had-crt2 This study
Plasmids
pEE32 pUC18 derivative; phaPCJAc [10]
pBPP pBBR ori (broad host range), mob, Kanr, PphaP1 [44]
pBPP-ccrMeJ4a-emd pBPP derivative; ccrMe, phaJ4a, emdMm [23]
pBPP-ccrMeJAc-emd pBPP derivative; ccrMe, phaJAc, emdMm This study
pColdII ColE1 ori, Ampr, PcspA Takara Bio
pColdII-phaB1 pColdII derivative, phaB1 This study
pColdII-phaB2 pColdII derivative, phaB2 This study
pET15b-phaB3 pET15b derivative, phaB3 This study
pColdII-phaB1NV pColdII derivative, phaB1NV This study
pColdII-phaB1NVYF pColdII derivative, phaB1NVYF This study
pK18mobsacB-AR pK18mobsacB derivative; phaB1 del [33]
pK18mobsacB-ABNVYFR pK18mobsacB-AR derivative; phaB1NVYF ins This study
pK18mobsacB-ABNVR pK18mobsacB-AR derivative; phaB1NV ins This study
pK18mobsacB-AB2R pK18mobsacB-AR derivative; phaB2 ins This study
pK18mobsacB-AHCR pK18mobsacB-AR derivative; had-crt2 ins This study
  1. The postfix del and ins indicate constructs for targeted gene deletion and insertion, respectively
  2. Ac, Aeromonas caviae; Me, Methylorubrum extorquens; Mm, Mus musculus. phaCNSDG, a gene encoding N149S/D171G mutant of PHA synthase from A. caviae. phaB1NV and phaB1NVYF, genes encoding N142V and N142V/Y185F mutants of NADPH-acetoacetyl-CoA reductase PhaB1 from R. eutropha, respectively