Table 1 Primers used in the study (restriction sites represented in upper case)
From: Enhancement of cellulosome-mediated deconstruction of cellulose by improving enzyme thermostability
Construct name (intermediate construct) | Vector | Forward primer (5’–3’) | Reverse Primer (5’–3’) | Linearized vector | Primary PCR templatea | Secondary PCR templatea |
|---|---|---|---|---|---|---|
BglA-v (BglA) | pET21a | CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga | ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc | pET28a BglA [32] | pET21a | |
BglA-v | pET21a | ttacatACTAGTtctcataaatttatctatgg | ttactaGAGCTCctattgttcttcaactggga | pET9d Xyn-docB [48] | – | |
BglA*-v (BglA*) | pET21a | CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga | ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc | pET28a BglA* | pET21a | |
BglA*-f | pET21a | CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga | ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc | pET28a BglA* | pET21a Xyn10A-f [42] | |
BglA*-v | pET21a | ttacatACTAGTtctcataaatttatctatgg | ttactaGAGCTCctattgttcttcaactggga | pET9d Xyn-docB [48] | – | |
8A-g (8A) | pET28a | ttacgtCCATGGgtgtgccttttaacacaaaata | ttacgaGGTACCaatgaaggtgtcggattcga | pET28a Cel8A [29] | – | |
8A-g | pET28a | tactagGGTACCagaagaagcaaacaagggag | ttactaCTCGAGcttacccagtaagccattct | pET28a-g-5A [6] | – | |
8A*-g (8A*) | pET28a | ttacgtCCATGGgtgtgccttttaacacaaaata | ttacgaGGTACCaatgaaggtgtcggattcga | pET28a 8AQM [24] | – | |
8A*-g | pET28a | tactagGGTACCagaagaagcaaacaagggag | ttactaCTCGAGcttacccagtaagccattct | pET28a-g-5A [6] | – | |
8A*-b | pET28a | ttacgtCCATGGgtgtgccttttaacacaaaata | ttacgaGGTACCaatgaaggtgtcggattcga | pET28a 8A-b [29] | ||
Scaf·GT | pET9d | GTTTAACTTTAAGAAGGAGATATAccatggtgcttcctccgaaaactacc | ggtgtgtttgtcggtgtgtttgtcGGTACCgcttcttcctgagagacaatc | pET28a CBM-Coh2375 [34] | pET28a Scaf·AT [49] | |
Scaf·GTV | pET9d | gttctttgacggtggagtaaatgttggagatacaacagtacctacaacacctacaacacctGCAGGGCAATTACAAATTGA | CCTTTCGGGCTTTGTTAGCAGCCggatcctAATTTGAGCCAACCAATATAG | pET28a CBM-CohC1 [48] | pET9d Scaf·GT |