Table 2 Bacterial strains and plasmids used in this study
From: Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant Escherichia coli
Name | Relevant characteristics | Source or reference |
|---|---|---|
Strains | ||
E. coli JM109 | recA1, endA1, gyrA96, thi, hsdR17, supE44, relA1, Δ(lac proAB)/F’ | TaKaRa (Dalian, China) |
[traD36, proAB+, lacqlacZΔ M15] | ||
E. coli JM109SG | JM109 ∆sad ∆gabD | [28] |
E. coli Trans1-T1 | The fastest growing chemically competent strain currently available | TransGen Biotech (Beijing, China) |
Ralstonia eutropha H16 | Wild type | ATCC17699a[43] |
Plasmids | ||
pKSSE5.3 | pBluescript vector derived, containing phaC and orfZ, AmpR | [22] |
pKSSEP1 | phaP1 gene inserted into pKSSE5.3, AmpR | This study |
pKSSEP2 | phaP2 gene inserted into pKSSE5.3, AmpR | This study |
pKSSEP3 | phaP3 gene inserted into pKSSE5.3, AmpR | This study |
pKSSEP4 | phaP4 gene inserted into pKSSE5.3, AmpR | This study |
pMCSH5 | sucD-4hbD inserted into pBBR1MCS-2, KmR | [28] |
Primers (5 ′ → 3′) | ||
phaP1F | AGTCTAGGCCT AAGAAATGCGCCTTGACCCACCC | This study |
phaP1R | AGTCTAGGCCT GCAAAACACACCGCAAACGCCAG | |
phaP2F | CAGCGAGGCCT GTTCGCAATGCTGCAATCTTTATT | This study |
phaP2R | ACTATAGGCCT ATACCACCCGTGACAACGGCAAG | |
phaP3F | ACTATAGGCCT GATTCGCACTCGGATGCTGCGCT | This study |
phaP3R | CAGCGAGGCCT TTGTATACCGATGCGGGAAGATT | |
phaP4F | CAGCGGACGTTGTC TCACGATGCAGCAATTGTTTTCC | This study |
phaP4R | AGTCTGACGTTGTC CTTCGACACGAAGGAAGTTTAGGC | |