Table 1 Bacterial strains and plasmids used in this study
From: Gene cloning and characterization of a novel esterase from activated sludge metagenome
Strain or plasmid | Description | Source or reference |
|---|---|---|
Strains | ||
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 TOP10 | lac x74 recA1 deoR F - mcrA Δ (mrr-hsdRMS-mcrBC) φ80 lacZ ΔM15 Δ araD139 Δ (ara-leu)7697 galU galK | TianGen |
E. coli BL21(DE3) | F-, ompT, hsdSB (rB-, mB-), dcm, gal, λ(DE3), pLysS, Cmr | Novogen |
E. coli EPI300-FosB12L1 | Positive clone from Fosmid genomic library, which carries the lipolytic gene | This study |
E. coli TOP10-EstAS | Positive clone from sublibrary, which carries the EstAS gene fragment | This study |
E. coli BL21(DE3)-EstAS | Positive clone, which carries the pEstAS-His expression vector | This study |
Plasmids | ||
pCC2FOS | Cloning vector; Cmr | Epicentre |
pUC118 | Cloning vector; Apr | Takara |
pET28a | Expression vector; Kmr | Novagen |
FosB12L1 | pCC2FOS, which carries the EstAS gene cluster (35 kb) | This study |
EstAS | pUC118, which carries the complete lipolytic gene (EstAS) | This study |
pEstAS-His | pET28a carrying amplified Hin dIII -Nde I fragment containing lipolytic gene (EstAS) | This study |