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Table 2 Strains, plasmids, and oligonucleotides used in this study

From: Secretion of polyhydroxybutyrate in Escherichia coli using a synthetic biological engineering approach

Strains Relevant characteristics Reference
Strains   
BL21-Gold (DE3) E. coli B F ompT hsdS(rB mB) dcm+ TetR gal λ(DE3) endA Hte Agilent technologies
XL1 Blue recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F’ proAB lacIqZΔM15 Tn10 (Tetr)] Agilent technologies
Cupriavidus necator H16 Wild type, PHA producing ATCC 17699
Plasmids   
pLG575 pACYC184 derivative, HlyBD, p15A origin, CmR [47]
pBHR68 pBluescript SK-, phbCAB genes from R. eutropha [16]
pSB1AK3 High copy BioBrick vector, pMB1 origin, AmpR and KanR [49]
pSB1A3 High copy BioBrick vector, pMB1 origin, AmpR [49]
pSB3K3 Medium copy BioBrick standard vector, p15A origin, KanR [49]
pCMEL1 phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pSB1A3 This study
pCMEL2 phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pSB3K3 This study
pCMEL3 phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pBHR68 This study
Oligonucleotides   
PhaP1FOR 5′- gaattcgcggccgcttctagaatgatcctcaccccggaaca-3 This study
PhaP1REV 5′- ctgcagcggccgctactagttcaggcagccgtcgtcttct-3′ This study
g114t 5′- cgtcgagctgaaccttcaggtcgtcaagact-3′ This study
g114t_antisense 5′-agtcttgacgacctgaaggttcagctcgacg-3′ This study