Table 1 Bacterial strains, plasmids and primers used in this study
Strain/plasmid/primer | Relevant characteristics | Reference/source |
|---|---|---|
Bacterial strains | ||
B. aryabhattai PHB10 | PHB accumulating environmental isolate | Pillai et al. (2017a), MTCC Accession no. 12561 |
E. coli JM109 | endA1, glnV44, recA1, gyrA96, thi, hsdR17 (r −k , m +k ), relA1, supE44, Δ(lac-proAB), [F′ traD36, proAB, laqIqZΔM15] | Promega Corporation |
Plasmids | ||
pUC18 | Cloning vector; AmpR | Thermo Fisher Scientific |
pGEM-T easy | T/A cloning vector; AmpR | Promega Corporation |
pTZ57R/T | T/A cloning vector; AmpR | Thermo Fisher Scientific |
pHB4485 | pUC18 carrying 4.48 kb PHA gene cluster from B. aryabhattai PHB10 | This work |
pHB5803 | pUC18 carrying 4.48 kb PHA gene cluster and phaA from B. aryabhattai PHB10 | This work |
Primers | ||
PHA4485F | 5′-GTTACCCCAAATTCTTGAGC-3′ | This work |
PHA4485R | 5′-CAGGAGTCTTCGCCTTGC-3′ | This work |
PHA1278F | 5′-GAAAGGAAATTGAGCAAGCG-3′ | This work |
PHA1278R | 5′-TGCTCCAATAACCATAACTG-3′ | This work |
T7 promoter primer | 5′-TAATACGACTCACTATAGGG-3′ | Promega Corporation |
SP6 promoter primer | 5′-TATTTAGGTGACACTATAG-3′ | Promega Corporation |
M13/pUC sequencing primer | 5′-GCCAGGGTTTTCCCAGTCACGA-3′ | Thermo Fisher Scientific |
M13/pUC reverse sequencing primer | 5′-GAGCGGATAACAATTTCACACAGG-3′ | Thermo Fisher Scientific |