Table 2 Strains, plasmids and primers used in this study
From: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy
Strain, plasmid, or primer | Relevant characteristics | Source or reference |
|---|---|---|
Strains | ||
 E. coli | ||
  DH5α | supE44 ΔlacU169(φ80 lacZΔM15) recA1 endA1 hsdR17(r _K m +K ) thi-1 gyrA relA1 F– Δ(lacZYA-argF) | Takara |
  BL21 (DE3) | F− ompT hsdS B (r −B m −B ) gal dcm (DE3) | Novagen |
 B. subtilis | ||
  168 | trpC2 | (Jongbloed et al. [15]) |
  Total-tat 2 | trpC2; tatAc::Em; tatAy-tatCy::Sp; tatAd-tatCd::Km; Emr; Spr; Kmr | (Jongbloed et al. [15]) |
  tatAyCy | trpC2; tatAy-tatCy::Sp; Spr | (Jongbloed et al. [15]) |
  tatAdCd | trpC2; tatAd-tatCd::Km; Kmr | (Jongbloed et al. [15]) |
Plasmids | ||
 pMD19-T | Apr, TA cloning vector for PCR product sequencing | Takara |
 pET30a | Expression vector, T7 promoter, Kmr | Novagen |
 pWH1520 | E. coli-B. subtilis shuttle vector, pBC16 ori, pBR ori, P xylA , xylR, Tcr, Apr | Mo Bi Tec |
 pET-c36 | pET30a derivative for expressing recombinant His6-tagged cel36 | This study |
 pET-c3623 | pET30a derivative for expressing recombinant His6-tagged cel3623 | This study |
 pET-c7482 | pET30a derivative for expressing recombinant His6-tagged cel7482 | This study |
 pWYC7 | pWH1520 derivative, secretion expression vector coding for YwbN-cel7482 fusion protein | This study |
Primers | ||
 F36c | 5′-ATGAAAAAATACCGCTGTAA-3′ | This study |
 R36c | 5′-TCACTTTTTCTCAGCGGCCA-3′ | This study |
 F3623c | 5′-ATGACAAATTTTACAGGCTT-3′ | This study |
 R3623c | 5′-TCATATTTTTCTGCTTACTA-3′ | This study |
 F7482c | 5′-ATGAAGAAATTTGACGGTTT-3′ | This study |
 R7482c | 5′-TCACTTTCTGCTTACAATCT-3′ | This study |
 F36e | 5′-CATATGAAAAAATACCGCTGTAAAAAGAAGGGAATTCTTTGGACACTGG-3′ | This study |
 R36e | 5′-CTCGAGTCAGTGGTGGTGGTGGTGGTGCTTTTTCTCAGCGGCCAGCCATATCTCGTCCA-3′ | This study |
 F3623e | 5′-CATATGACAAATTTTACAGGCTTCAGTGCAGGAGTAAATCTTGGAGGC-3′ | This study |
 R3623e | 5′-CTCGAGTCAGTGGTGGTGGTGGTGGTGTATTTTTCTGCTTACTATCTTAATCAAATCCTCAT-3′ | This study |
 F7482e | 5′-CATATGAAGAAATTTGACGGTTTCATGGCGGGAGTAAACCTTGGG-3′ | This study |
 R7482e | 5′-CTCGAGTCAGTGGTGGTGGTGGTGGTGCTTTCTGCTTACAATCTTAACCAGTTCGTC-3′ | This study |