Table 1 Strains, plasmids, and primers used in this study
From: Direct cadaverine production from cellobiose using β-glucosidase displaying Escherichia coli
Strain, plasmid, or primer | Relevant phenotype, description, or sequence (5′-3′) | Source or reference |
|---|---|---|
Strains | ||
Escherichia coli | ||
NovaBlue | endA1 hsdR17 (rK12- mK12+) supE44 thi-1 recA1 gyrA96 relA1 lac[F’ proAB+ lacIqZΔM15::Tn10 (Tetr)]; host for DNA manipulation | Novagen |
JCM20137 | National Institute of Genetics, Japan | |
Japan Collection of Microorganisms, RIKEN BRC | ||
Jm-cadA | JCM20137 strain harboring pHLA-cadA vector | This study |
Jm-blc-Tfu | JCM20137 strain harboring pHLA-blc-Tfu vector | This study |
Jm-cadA-blc-Tfu | JCM20137 strain harboring pHLA-cadA-blc-Tfu vector | This study |
Genomic DNA | ||
Thermobifida fusca YX | ATCC BAA-629D-5 | ATCC |
Plasmids | ||
pHLA | Cell surface display vector containing pgsA gene under HCE promoter control, ampicillin resistance marker | Tanaka et al. 2011 |
pHLA-cadA | Vector for CadA expression | This study |
pHLA-blc-Tfu | Vector for BGL (Tfu0937 from T. fusca) expression using blc anchor protein | Tanaka et al. 2011 |
pHLA-cadA-blc-Tfu | Vector for CadA and BGL (Tfu0937 from T. fusca) coexpression using blc anchor protein | This study |
Oligonucleotide primers | ||
CadA_F | agcagatctgatgaacgttattgcaatattgaatcacatggggg | |
CadA-FLAG_R | ccagtcgacttatttatcgtcatcatctttataatcttttttgctttcttc | |
HCE-blc-Tfu-term_F | cgccgtagcgccgatggtagtgtggggtctccccatgcgag | |
HCE-blc-Tfu-term_R | ggagagatcgcggccgccatggggtcaggtgggaccaccgcgctac | |
pHLA-cadA_F | gcggccgcgatctctccttcacagattcccaatctcttgtt | |
pHLA-cadA_R | ctcgcatggggagaccccacactaccatcggcgctacggcg |