Table 1.
Starting bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| E. coli TOP10 | lacх74 recA1 deoR F - mcrA ∆ (mrr-hsdRMS-mcrBC) ϕ80 lacZ∆M15∆ araD139∆ (ara-leu)7697 galU galK | Transgen |
| E. coli EPI300™-T1R | [F- e14-(McrA-) D(mcrC-mrr) (TetR) hsdR514 supE44 supF58 lacY1 or D(lacIZY)6 galK2 galT22 metB1 trpR55 l-] | Epicentre |
| E. coli BL21(DE3) | F-, ompT, hsdSB (rB-, mB-), dcm, gal, λ(DE3), pLysS, Cmr | Novogen |
| E. coli EPI300-FosD11L2 | Positive clone from Fosmid genomic library, which carries the lipolytic gene | This study |
| E. coli TOP10-EstMY | Positive clone from sublibrary, which carries the EstMY gene fragment | This study |
| E. coli BL21(DE3)-EstMY | Positive clone, which carries the pEstMY-His expression vector | This study |
| Plasmids | ||
| pCC1FOS | Cloning vector; Chlr | Epicentre |
| pUC18 | Cloning vector; Apr | Takara |
| pET28a | Expression vector; Kmr | Novagen |
| FosD11L2 | pCC1FOS, which carries the estMY gene cluster (31 kb) | This study |
| pUC18-EstMY | pUC18, which carries the complete lipolytic gene (estMY) | This study |
| pEstMY-His | pET28a carrying amplified HindIII -NdeI fragment containing lipolytic gene (estMY) | This study |
Apr, ampicillin resistant; Chlr, chloramphenicol resistant; Kmr, kanamycin resistant.