TABLE 1.
Primers used to clone the genes in this study
Mutation points are shown in bold.
| Genes | Residues | Primers | Direction |
|---|---|---|---|
| BhCel5B | 26–574 | GGATCCGTTAGTTCTGCTCATGAGGATGTG | Forward |
| GTCGACATTCGGGTAACACCATAGAAAGC | Reverse | ||
| BhCBM46 | 457–574 | TGGGATCCTATCGTACGCCTGTATTGC | Forward |
| GTCGACGGGTAACACCATAGAAAGCGCTT | Reverse | ||
| BhGH5-Ig | 30–463 | CACACGCTAGCGCTCATGAGGATGTGAAAAC | Forward |
| CACACCTCGAGTTGCAATACAGGCGTACG | Reverse | ||
| BhIg-CBM46 | 365–574 | CACACGCTAGCTCCGTTGCCGAGTCAAAC | Forward |
| CACACCTCGAGTGCGGCCGCAAGCTTGTCG | Reverse | ||
| BhCel5B_E296A | 26–574 | GGAATTCCAGTCGTTCTAGGT[b]GCGTTCGGCTTGCTTGGATTTG | Forward |
| CAAATCCAAGCAAGCCGAA[b]CGCACCTAGAACGACTGGAATTCC | Reverse | ||
| BhGH5-Ig_E296A | 30–463 | CACACGCTAGCGCTCATGAGGATGTGAAAAC | Forward |
| CACACCTCGAGTTGCAATACAGGCGTACG | Reverse | ||
| BhCel5B_W501A_E296A | 26–574 | GGAAATGCTGGCCCGCAAGAC[b]GCTACTTCCTTTAAGGAGTTTGG | Forward |
| CCAAACTCCTTAAAGGAAGT[b]AGCGTCTTGCGGGCCAGCATTTCC | Reverse | ||
| BhCel5B_F504A_E296A | 26–574 | GGCCCGCAAGACTGGACTTCC[b]GCCAAGGAGTTTGGCTATGCC | Forward |
| GGCATAGCCAAACTCCTT[b]GGCGGAAGTCCAGTCTTGCGGGCC | Reverse | ||
| BhCel5B_F507A_E296A | 26–574 | GACTGGACTTCCTTTAAGGAG[b]GCCGGCTATGCCTTCTCTCCTTC | Forward |
| GAAGGAGAGAAGGCATAGCC[b]GGCCTCCTTAAAGGAAGTCCAGTC | Reverse | ||
| BhCel5B_Y509A_E296A | 26–574 | CCTTTAAGGAGTTTGGC[b]GCCGCCTTCTCTCCTTCATATGATGC | Forward |
| GCATCATATGAAGGAGAGAAGGC[b]GGCGCCAAACTCCTTAAAGG | Reverse | ||
| BhCel5B_R531A_E296A | 26–574 | GGCGTTTTTTCGTGAGGTG[b]GCCGATGGTGAAGTTCGGTTAACC | Forward |
| GGTTAACCGAACTTCACCATC[b]GGCCACCTCACGAAAAAACGCC | Reverse | ||
| BhCel5B_W501A_F504A_F507A_Y509A_R531A_E296A | 30–574 | Gene synthesized | |