Table 2 Bacterial strains and plasmids
From: Metabolic engineering of Agrobacterium sp. strain ATCC 31749 for production of an α-Gal epitope
Strains or plasmids | Description | Source |
|---|---|---|
ATCC 31749 | Curdlan-producing Agrobacterium sp. | ATCC |
LTU265 | ATCC 31749 locus II mutant with decreased curdlan production | B.A. Stone V.A. Stanisich |
JM109 | E. coli K12 strain used for lacY cloning | Promega |
ATCC 31749ΔcrdS | ATCC 31749 with interrupted curdlan synthase gene (crdS) | This study |
pGEM-T easy | Vector used for gene cloning | Promega |
pBQ | Broad-host-range expression vector constructed for gene expression in ATCC 31749 | [15] |
pET15b-αGalT | Plasmid containing truncated bovine α1,3-galactosyltransferase gene | [11] |
pT-α1,3-galT | pGEM-T easy vector containing truncated bovine α1,3-galactosyltransferase with BamHI and SacI restriction sites | This study |
pT-GalE | Plasmid containing UDP-galactose 4'-epimerase (galE) from E. coli with a short linker sequence for fusion enzyme construction | [15] |
pTET | pGEM-T easy vector containing the gale:α1,3galT fusion gene | This study |
pBQET | Plasmid for GalE:α1,3GalT fusion enzyme expression in ATCC 31749 and ATCC 31749ΔcrdS | This study |
pT-lacY | pGEM-T easy vector containing lactose permease gene (lacY) from JM109 with SacI and XhoI restriction sites | This study |
pBQETY | Plasmid for expression of GalE:α1,3GalT fusion enzyme and E. coli lactose permease (LacY) in ATCC 31749 and ATCC 31749ΔcrdS | This study |
pT-crdS | pGEM-T easy vector containing curdlan synthase gene (crdS) with SacI and KpnI restriction sites | This study |
pT-crdSh | pGEM-T easy vector containing two 500 bp regions of homology to crdS with SacI/KpnI and EcoRI/HindIII restriction sites | This study |
pBluescript II (KS-) | Phagemid for gene knockout in ATCC 31749 | Stratagene |
pBScrdSh | pBluescript II (KS-) containing two 500 bp regions of homology to crdS with EcoRI and HindIII restriction sites | This study |
pYanni2 | Plasmid containing a gentamicin resistance cassette | [28] |
pT-GmR | pGEM-T easy vector containing gentamicin resistance cassette with EcoRI and HindIII restriction sites | This study |
pBScrdShG | crdS knockout plasmid; pBluescript II (KS-) containing a gentamicin resistance cassette flanked by two 500 bp regions of homology to crdS | This study |