Table 3 The strains, plasmids and primers used in this study
From: Construction of cellulose-utilizing Escherichia coli based on a secretable cellulase
Description | Description | Source |
|---|---|---|
Strains | ||
E. coli DH5α | F− endA1 hsdR17(r K − m K +) supE44 thi-l λ − recA1 gyrA96 ΔlacU169 (Φ80dlacZM15) | Invitrogen |
E. coli BL21(DE3) | hsdS, gal(ΔcIts857 ind1, Sam7, nin5, lacUV5-T7, gene1 | Novagen |
Plasmids | ||
pACYCDuet | 4.0 kb, p15A ori, T7 promoter, CmR | Novagen |
pACYCDuet/cel | pACYCDuet, cel-cd gene under T7-1 promoter | This study |
pACYCDuet/n20 | pACYCDuet, n20 gene under T7-1 promoter | This study |
pCel-Tfu | cel-cd gene fusion with tfu from Thermobifida fusca | This study |
pN20-Tfu | n20 gene fusion with tfu from Thermobifida fusca | This study |
pBHR68 | pSK-derivative, phbA, phbB and phbC from Ralstroniaeutropha | [31] |
pN20XynC-A | n20 gene fusion with xynC from F. succinogenes | This study |
pN20Xyn10B | n20 gene fusion with xyn10b from C. stercorarium | This study |
pN20PelC | n20 gene fusion with pelC from B. subtilis | This study |
pCelPelC | cel-cd gene fusion with pelC from B. subtilis | This study |
pCelXsa | cel-cd gene fusion with xsa from B. ovatus | This study |
Primers | Sequences |
|---|---|
Cel-NcoI-F | 5′- TTTTCCATGGAAGGAAACACTCGTGAAGA-3′ |
Cel-BamHI-R | 5′- TTTTGGATCCAAGTACTTTCGTGTATTTTG-3′ |
Cel-20-BamHI-R | 5′- TTTTGGATCCGCGTTTAACATTGTCATTAC-3′ |
Tfu-BamHI-F | 5′-TTTTGGATCCACCTCGCAATCAACGACGCC-3′ |
Tfu-XhoI-R | 5′-TTTTCTCGAGTTATTCTTGACCGAAAATACCAC-3′ |
XynC-A-BamHI-F | 5′- TTTTGGATCC CAAGACTTCTGCAGCAACGC-3′ |
XynC-A-XhoI-R | 5′-TTTTCTCGAGGTTTTTAACATAAACTTTCG-3′ |
Xyn10B-BamHI-F | 5′- TTTTGGATCCAACAAGTTCCTGAACAAGAA-3′ |
Xyn10B-XhoI-R | 5′-TTTTCTCGAGTTCGCGCAGACGAGACGGAT-3′ |
PelC-BamHI-F | 5′- TTTTGGATCCGCAGATAAGGTTGTCCACGA-3′ |
PelC-XhoI-R | 5′- TTTTCTCGAGTTAGAACTGGGTGTTATTAT-3′ |
Xsa-BamHI-F | 5′- TTTTGGATCCAAGACCGAAAAGCGTTACCT-3′ |
Xsa-XhoI-R | 5′-TTTTCTCGAGTTCGTCTTTGCCTTCAATGG-3′ |